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Sponges (phylum Porifera) are ancient, filtering, multicellular metazoans. Freshwater sponges (Demosponges, Lubomirskiidae) dominate the fauna of the littoral zone of Lake Baikal. Over the last years, there have been mass diseases and death of endemic sponges. Previously, the strain Janthinobacterium sp. SLB01 was isolated from the diseased sponge Lubomirskia baicalensis. The studies of the pathogenicity of the strain Janthinobacterium sp. SLB01 for Baikal sponges has not been carried out. Therefore we infected experimentally in vitro to determine its pathogenicity by the cell culture of the primmorphs with subsequent isolation, sequencing, and analysis of the genomes. The purpose of the study is to show that the strain Janthinobacterium sp. SLB01 isolated from the diseased sponge L. baicalensis is a pathogen for the cell culture of primmorphs. The bacteria from the infected samples of the primmorphs were isolated and identified as strain Janthinobacterium sp. PLB02. A comparative analysis of the genomes of the strains Janthinobacterium sp. SLB01 and Janthinobacterium sp. PLB02 showed that they are practically identical. The genomes of both strains contain genes vioABCDE violacein, flok formation, and strong biofilm, and the type VI secretion system (T6SS), as the primary virulence factor. The bacterial strains Janthinobacterium sp. based on a comparison of complete genomes showed similarity with strain Janthinobacterium lividum MTR. Isolated our strains of Janthinobacterium sp. are pathogens for cell cultures of primmorphs and L. baicalensis sponges. The results of the study will help to expand the understanding of microbial relationships in the development of disease and the death of Baikal sponges.

The methods for data presentation are important in bioinformatics as data processing algorithms. The article describes the software package for the extensive analysis of tables with estimates of bacterial abundance levels in environmental samples. The package was designed to be executed in a distributed hardware environment, with powerful packages in Python in the backend and interactive front-end forms. Most of microbial ecology-specific functionality is implemented by the scikit-bio Python package, together with the other Python packages intended for big data analysis. Interactive visualisation tools are implemented by the D3.js software library, therefore, the software project is named D3b. The package is a suite of tools for the analysis of microbial ecology data implemented as a web-service and as a desktop application. It supports a substantial part of the graphical and analytical descriptions of microbial communities used in scientific publications. Source codes are available at github (sferanchuk/d3b_charts) and the on-line version of the system is accessible at d3b-charts.bri-shur.com.

The complete mitochondrial genome of the Lake Baikal sponge Baikalospongia intermedia was sequenced. The circular mitochondrial genome is 28,327 bp in length and includes 14 protein-coding genes, 2 ribosomal RNA genes and 25 transfer RNA genes. Bayesian comparative analysis of molecular evolution rates was found no acceleration of the mtDNA evolution of B. intermedia. This species clustered with other species of the genus Baikalospongia on the Bayesian tree.

Endemic freshwater sponges (demosponges, Lubomirskiidae) dominate in Lake Baikal, Central Siberia, Russia. These sponges are multicellular filter-feeding animals that represent a complex consortium of many species of eukaryotes and prokaryotes. In recent years, mass disease and death of Lubomirskia baicalensis has been a significant problem in Lake Baikal. The etiology and ecology of these events remain unknown. Bacteria from the families Flavobacteriaceae and Oxalobacteraceae dominate the microbiomes of diseased sponges. Both species are opportunistic pathogens common in freshwater ecosystems. The aim of our study was to analyze the genomes of strains Janthinobacterium sp. SLB01 and Flavobacterium sp. SLB02, isolated from diseased sponges to identify the reasons for their joint dominance. Janthinobacterium sp. SLB01 attacks other cells using a type VI secretion system and suppresses gram-positive bacteria with violacein, and regulates its own activity via quorum sensing. It produces floc and strong biofilm by exopolysaccharide biosynthesis and PEP-CTERM/XrtA protein expression. Flavobacterium sp. SLB02 utilizes the fragments of cell walls produced by polysaccharides. These two strains have a marked difference in carbohydrate acquisition. We described a possible means of joint occupation of the ecological niche in the freshwater sponge microbial community. This study expands the understanding of the symbiotic relationship of microorganisms with freshwater Baikal sponges.

Green algae of the phylum Chlorophyta are the most widespread autotrophic picoplankton in Lake Baikal (Russia). To expand our molecular biological knowledge of these microalgae and compare them in the future with an endosymbiotic strain, we present here the draft genome sequence of Chlorella sp. strain BAC9706., Green algae of the phylum Chlorophyta are the most widespread autotrophic picoplankton in Lake Baika (Russia). To expand our molecular biological knowledge of these microalgae and compare them in the future with an endosymbiotic strain, we present here the draft genome sequence of Chlorella sp. strain BAC9706.

Green algae of the phylum Chlorophyta are the most widespread autotrophic picoplankton in Lake Baikal (Russia). To expand our molecular biological knowledge of these microalgae and compare them in the future with an endosymbiotic strain, we present here the draft genome sequence of Chlorella sp. strain BAC9706.

Endemic freshwater sponges (Demosponges, Lubomirskiidae) dominate in Lake Baikal and are multicellular filter-feeding animals represent a complex consortium of many species of eukaryotes and prokaryotes. In recent years, mass disease and death of the L. baicalensis have been an urgent problem of Lake Baikal. The etiology and ecology of these events remain unknown. Bacteria in microbiomes of diseased sponges of the families Flavobacteriaceae and Oxalobacteraceae were dominant. Both species are opportunistic pathogens common for freshwater ecosystems. The aim of our study is to analyze the genomes of strains Janthinobacterium sp. SLB01 and Flavobacterium sp. SLB02, isolated from diseased sponges to identify the reasons for their joint dominance. The first one attacks the other cells using type VI secretion system, suppress gram-positive bacteria with violacein pigment and regulate its own activity via quorum sensing. It makes the floc and strong biofilm by exopolysaccharide biosynthesis and PEP‐CTERM proteins expression. The second one utilizes the fragments of cell walls produced of polysaccharides. Named two strains have noticeable difference in carbohydrates acquisition. We described the possible way of joint occupation of ecological niche into freshwater sponge microbial community. This study expands understanding about symbiotic relationship of microorganisms with freshwater Baikal sponges.

There are significant changes in the consortium of microorganisms of freshwater Baikal sponges during their mass death, which began in 2011. The alleged cause of disease is a significant increase in the number of opportunistic microorganisms. Here, we report the draft genome sequence of Flavobacterium sp. strain SLB02.

Background. Freshwater sponges (Demosponges, Lubomirskiidae) are dominated in the littoral zone Lake Baikal in the biomass of benthic organisms and represent complex consortia of many species of eukaryotes and prokaryotes. A distinctive feature of sponges from Lake Baikal is their ability to live in symbiosis with various kinds of chlorophyll containing microalgae. Recently there have been massive diseases and the death of freshwater sponges. The etiology and ecology of these events remain unknown. Purpose. The purpose of the research was to use cell culture of primmorphs in vitro to study the microbiomes of healthy and diseased sponges to show the transmission of pathogenic agents from diseased sponges to cell cultures. Methods. The cell culture of primmorphs sponge Lubomirskia baicalensis was used to study microbiome communities in diseased and sick sponges in comparison with healthy sponge with subsequent sequencing of gene 16S rRNA and analysis of changes in microbiomes. Results. Results this study were show that use of cell culture of primmorphs in vitro is equivale of healthy sponge. Microbial community of healthy sponge and primmorphs was grouped separately from the community of diseased sponges and infected primmorphs, which confirms the suitability the cell culture of primmorphs, as a model sponge system. We found the mass death of green symbionts (Chlorophyta) and a shift in the microbial communities of sponges/primmorphs, associated with increase in relative abundant of different phyla Bacteroidetes and Proteobacteria with dominated families Flavobacteriaceae and Burkholderiaceae, Moraxellaceae in diseased sponges and infected cell cultures of primmorphs. Conclusions. This approach allowed us, using the cell culture of primmorphs, to identify potential opportunistic bacteria that can work together, which possibly enhances their action. The primmorphs system described here is a powerful new model system for studying basic mechanisms of the development of sponge disease, which will be valuable in future studies.

Sponges are an ecologically important component of marine and freshwater bodies. Sponge community includes a variety of microorganisms: fungi, algae, archaea, bacteria and viruses. Despite active research in the field of aquatic virology, biodiversity and the role of viruses in sponges are poorly studied. The relevance of research in this area is also related to the worldwide problem of sponge diseases. The aim of this study was to elucidate the genetic diversity of viruses in the associated community of diseased endemic Baikal sponge Lubomirskia baikalensis using metagenomic analysis. As a result, we have shown for the first time a high genetic and taxonomic diversity of DNA viruses in the Baikal sponge community. Identified sequences belonged to 16 viral families that infect a wide range of organisms. Moreover, our analysis indicated the differences in viral communities of visually healthy and diseased branches of the sponge. The approach used in this study is promising for further studies of viral communities in sponges, obtaining more complete information about the taxonomic and functional diversity of viruses in holobionts and entire Lake Baikal, and identifying the role of viruses in sponge diseases.

The spatial distribution of healthy and diseased sponges in Lake Baikal was investigated. The endemic sponges were studied in May − June 2015 at 11 transects within the depth range of 0 to 40 m. More than 500 sponge specimens were collected, and 29 taxa including 12 species and 17 morphospecies (sp. taxa) were identified. Among the 29 taxa, only B. martinsoni specimens and 14 morphospecies were healthy, whereas specimens of the other 11 species and 3 morphospecies were diseased, making up 35% of the total number of specimens (512). The different body forms of sponge found among the specimens were encrusting (66%), globulous (21%), and branched (13%). Sponges with necrosis signs, discolored specimens or covered with violet film or brown spots, were found in all transects under study (37 stations of 68). The percent cover of the bottom (per m2) by various body forms of sponges was evaluated with a sponge faunal state assessment test. Changes in the diversity and abundance of sponges due to their disease may jeopardize the functioning of the entire ecosystem of Lake Baikal which is a World Heritage Site. This baseline data on patterns of sponges, including their spatial distribution and percent cover, will be utilized to support future management and monitoring of Lake Baikal. Data on the current state of Baikal endemic sponge fauna are essential for understanding the causes and prognosis of disease processes of freshwater sponges.

In recent years, signifcant changes in the ecological system of the coastal (littoral) zone, including mass death of the endemic representatives of the freshwater sponges of the Lubomirskiidae family, have been an urgent problem of Lake Baikal. Similar problems are known all over the world. Thus, mass disease and death of corals and sponges are indicated in the Mediterranean, Adriatic, Caribbean and other seas, which raises serious concerns about the future of these biocenoses (Olson et al., 2006; Webster, 2007; Wulff et al., 2007; Stabili et al., 2012). In Baikal, diseased sponges were frst found in 2011. The area of sponge disease is constantly expanding, and, to date, dying specimens have been found throughout the lake. The mass death of sponges occur in presence of the large-scale violation of the spatial distribution and the structure of phytocenoses in the littoral zone, but the causes of these phenomena are unknown. The relevance of the problem arises from the fact that changes in the littoral zone of Lake Baikal can signifcantly affect the productivity and composition of planktonic organisms and zoobenthos, which are the food base for fsh, as well as the quality of drinking water. The deterioration of the ecological state also affects the attractiveness of the lake for tourism. At the international level, serious intellectual and fnancial resources were mobilized to solve similar problems. Despite the obvious relevance, in Russia such studies are carried out irregularly by small groups of researchers.

A comparative study of biological, molecular and genetic characteristics of a collection of ten strains of tick-borne encephalitis virus (TBEV) isolated in Primorsky Krai before 1960 and stored in a lyophilized state for a prolonged period (over 65 years) is presented. The collection includes the Sofjin strain isolated from the brain of a fatal case in Primorsky Krai in 1937 and transferred to the Scientific Research Institute of Epidemiology and Microbiology (Vladivostok) in 1953. All lyophilized viral strains demonstrated great preservation and high infectious activity in the model of 2-day-old non-inbred mice. Whole-genome sequencing showed that all strains belong to the Far East TBEV subtype, comprising three clusters of Sofjin-, Oshima- and Senzhang-like strains. We show that SofjinPYB, Sofjin (Vector) and Sofjin-HO strains form a separate branch of the phylogenetic tree and are closely related to Khabarovsk-Obor-4, but not to the original Sofjin strain. The Sofjin-1953, Sofijin-Chumakov, SofjinKSY and SofjinCDC strains are genetically close to each other and can be used as reference strains for comparative analysis of the tick-borne encephalitis virus population.

The draft genome sequence of Janthinobacterium sp. strain SLB01, a violacein-producing psychrotolerant bacterium isolated from the diseased sponge Lubomirskia baicalensis, was determined. We identified five genes encoding VioA, VioB, VioC, VioD, and VioE proteins related to violacein biosynthesis that were like those identified in published Janthinobacterium lividum strains MTR and RIT308.

Molecular-genetic analysis of cyanophages of the family Myoviridae from the associated community of the endemic Baikal sponge Lubomirskia baicalensis was carried out based on the g20 gene fragment. A high cyanophage diversity according to the g20 marker gene was found in the sponge. The Baikal sponge cyanophages were shown to be similar to those inhabiting plankton. Moreover, specific cyanophage groups that are significantly different from all of the known groups inhabiting the Lake Baikal were revealed.

In the flavivirus genus, the non-structural protein NS5 plays a central role in RNA viral replication and constitutes a major target for drug discovery. One of the prime challenges in the study of NS5 protein is to investigate the interplay between the two protein domains, namely, the RNA-dependent RNA polymerase (RdRp) domain and the methyltransferase (MTase) domain. These investigations could clarify the multiple roles of NS5 protein in the virus life cycle. Here we present the results of sequence analyses and structural bioinformatics studies of NS5 protein, which suggest that the conserved motif F in the NS5 protein could act as a lock which controls the rearrangement of the domains and as a switch in the protein enzymatic activity.

Sponges are permanent benthos residents which establish complex associations with a variety of microorganisms that raise interest in the nature of sponge-symbionts interactions. A molecular approach, based on the identification of the 16S rRNA and ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit genes, was applied to investigate diversity and phylogeny of bacterial phototrophs associated with four species of Lubomirskiidae in Lake Baikal. The phylogeny inferred from both genes showed three main clusters of Synechococcus associated with Baikalian sponges. One of the clusters belonged to the cosmopolitan Synechococcus rubescens group and the two other were not related to any of the assigned phylogenetic groups but placed as sister clusters to S. rubescens. These results expanded the understanding of freshwater sponge-associated photoautotroph diversity and suggested that the three phylogenetic groups of Synechococcus are common photosynthetic symbionts in Lubomirskiidae sponges.

Complete and nearly complete mtDNA genome sequences were used to resolve differences between two palaeontology-based hypotheses on timing of the origin of the Baikal endemic sponge family Lubomirskiidae (Haplosclerida, Demospongiae). Bayesian ratio test, when coupled with estimates of substitution rates based on known palaeontological findings, provided strong evidence for the Miocene origin over the Late Oligocene one. The common ancestor of the present day sponges in Lake Baikal diverged about 2,3 million years ago (Ma), while Lubomirskia baicalensis, Rezinkovia echinata, Baikalospongia intermedia profundalis and B. bacillifera split in the Pleistocene about 0,7 Ma. A phylogenetic analysis within the family suggested that speciation may have coincided with the occurrence of cold climatic conditions. We argue that the cause of speciation may be niche splitting due to temperature, depth and possibly feeding preferences.

Dermacentor nuttalli is an epidemiologically important tick in Palearctic Asia which transmits several infectious diseases including tularemia, North Asian tick-borne rickettsiosis, Lyme disease and tick-borne encephalitis. The genetic specificity and phylogeny of D. nuttalli from four geographic localities in Eastern Siberia were characterized using the mitochondrial (mt) 16S ribosomal RNA (rRNA) gene and internal transcribed spacer 2 (ITS2). Low genetic diversity was observed in the populations of ticks distributed from South Siberia to North China. From 11 detected mt 16S haplotypes, one was found in all populations, whereas the others were restricted to specific localities. These results suggested that the genetic structure of D. nuttalli represents integrated populations with no geographic isolation across the distribution area. The phylogenetic reconstructions inferred from the mt 16S rRNA gene and ITS2 were in agreement and showed a distinct D. nuttalli clade within a monophyletic Eurasian lineage of Dermacentor sp.

The disease of freshwater sponges was first discovered in 2011, when pink samples were found in the Central Basin of Lake Baikal. Subsequently, the visible signs of the disease have changed, and now sponges appear with various symptoms of damage to the body, such as discoloration, tissue necrosis, the formation of brown patches and dirty-purple biofilms on some branches. These signs of the disease are accompanied by the mass death of sponges. We identified differences in microbiomes by sequencing 16S rRNA genes and found changes in the consortium of microorganisms of freshwater Baikal sponges. We found that the observed imbalance in the studied microbial communities of diseased sponges is caused by several different conditionally pathogenic microorganisms that increase their negative effect by acting together and in concert, which leads to the death of photosynthetic microalgae and sponges. Sponges are an important component of coastal communities, and the massive loss of sponges can obviously affect the structure of benthic communities and the purity of water.

Flavavirus RdRp contains two zinc atoms located in two specific zinc binding sites in the protein. Molecular dynamics experiments with flavavirus RdRp suggest that the conformation of the conservative motif F in the polymerase is highly sensitive to the bond lengths between the zinc atoms and their four coordinating atoms. In the experimental structures of flavavirus RdRp, motif F is presented in two different conformations. The polymerase acts as a catalyst only in one of the motif F conformations. We hypothesize that the second conformation is required for the formation of the viral replicative complex and that it is the quantum transition in the zinc atoms which slightly changes the bond lengths; thus assisting the switch between the two motif F conformations.

The NS3 protease and NS5 polymerase structures of the Zika virus were constructed and processed using a virtual screening pipeline. MM-PBSA calculations show that some of the ligands found by the pipeline demonstrate a good affinity to vulnerable sites in both proteins. The anti-hypertension drug eprosartan is among the selected ligands; and inhibition of the tick-borne encephalitis virus has already been confirmed in vivo by a previous study. In the present study phytochemicals bisabolol and andrographolide are suggested as the candidates for antiviral therapy.

Silicateins found in spicules of siliceous sponges are proteins that take part in biogenic silica precipitation and determine the morphological features of spicules. The exon-intron structure of the genes encoding four silicatein-α isoforms (−α1, −α2, −α3, and −α4) from an endemic Baikalian sponge Lubomirskia baicalensis was studied. For eight sponge species, including both cosmopolitan (Spongilla lacustris, Ephydatia muelleri, E. fluviatilis) and endemic Baikalian (L. baicalensis, L. incrustans, Baikalospongia intermedia, B. fungiformis, Sw. papyracea) species, seventeen partial sequences of different silicatein isoform genes were determined. It was shown that cosmopolitan and endemic Baikalian sponges differ from each other in gene structure, in particular, in intron length. Among Baikalian sponges, silicatein-α1 genes had the highest variation of intron length, and silicatein-α4 genes were the most conservative. A phylogenetic analysis based on amino acid sequences of different silicatein isoforms identified four distinct clusters within the freshwater sponge clade. An analysis based on exon-intron gene sequences enables discrimination between different sponge species within the clusters.

Morphological and molecular genetic data for freshwater sponges from the lakes of Tuva Depression, Baikalospongia dzhegatajensis (Rezvo, 1936), forms Dzh05 and Dzh06, from Chagatai Lake, as well as forms TKh1 and TKh2, from the Lake Tore-Khol, were obtained and examined. In the sponges examined, which on phylogenetic tree clustered together with the Ephydatia fluviatilis (Linneaus, 1758) sponge from the family Spongillidae, the ITS rDNA regions were sequenced. Comparison of highly variable interal spacer regions of the mitochondrial genome was performed using corresponding sequences of three sponges from the family Spongillidae (E. fluviatilis, E. muelleri and Spongilla lacustris), sponges from the Chagatai and Tore-Khol lakes (Dzh06 and TKh2) with an unknown status, and sponges from the Baikalian family Lubomirskiidae. Minimum genetic differences were observed between E. fluviatilis, Dzh06, and TKh2 (from 0.003 to 0.01% of nucleotide substitutions), while maximum differences were found between the species of Lubomirskiidae and Spongillidae (from 0.928 to 2.06%). The data obtained indicated that sponges from Chagatai and Tore-Khol lakes were most close to E. fluviatilis.

Background: The study of ecosystems of the great lakes is important as observations can be extended to ecosystems of larger scale. The ecological crisis of Lake Baikal needs investigations to discover the molecular mechanisms involved in the crisis. The disease of Baikal sponges is one of the processes resulting in the degradation of the littoral zone of the lake. Methods: The chloroplast genome fragment for the algae endosymbiont of Baikal sponge was assembled from metagenomic sequencing data. The distributions of polymorphic sites were obtained for the genome fragment, separately for samples from healthy sponge, diseased sponge and dead sponge tissues. Results: The comparative analysis of chloroplast genome sequences suggests that the symbiotic algae from Baikal sponge is close to Choricystis genus of unicellular algae. Also, the distributions of polymorphic sites allowed detection of the signs of extensive mutations in the chloroplasts isolated from the diseased sponge tissues. Conclusions: The study demonstrate the particular case of evolution at the molecular level due to the conditions of a severe crisis of a whole ecosystem in Lake Baikal. The detection of adaptive mutations in the chloroplast genome is an important feature which could represent the behavior of an ecosystem in the event of a severe crisis.

Genomes of four tick-borne encephalitis virus strains, isolated from the blood of the individuals after tick bites and causing no clinical symptoms of infection, were characterized. Analysis of translated polypeptides revealed 21 amino acid positions typical of this group of strains and distinguishing them from the other tick-borne encephalitis virus strains of Far Eastern subtype examined earlier. Only three mutations led to substantial amino acid changes, which probably could affect the infection process severity. It is suggested that two associated mutations, deletion of amino acid 111 in the capsid protein C and substitution (Ser1534 → Phe) in the NS3 protein influence strictly coordinated polyprotein processing, disturbing correct arrangement of viral particles. This process can result in the development of defect viral particles, containing no RNA. Mutation (Ser917 → Gly) in nonstructural protein NS1 results in the substitution of hydrophilic amino acid, specific to highly virulent strains, by the hydrophobic one. This could influence the effectiveness of viral replication complex, thereby affecting the infectivity of tick-borne encephalitis virus strains.

Siliceous spicules of Baikal freshwater sponge Lubomirskia baicalensis contain several proteins, including silicateins. Analysis of a L. baicalensis cDNA library revealed four different mRNAs coding for proteins related to marine sponge silicatein α (α1, α2, α3, and α4). The intron-exon structure was determined forthe genomic α1 silicatein gene. The gene is 1988 bp from the initiation to the termination codon and consists of six intron (total size 1007 bp) and seven exons (total size 981 bp). Mass spectrometry of a tryptic digest of spicule proteins revealed peptides of two silicateins α.

The phylogenetic relationships between freshwater sponges (families Lubomirskiidae, Spongillidae and Potamolepidae) are still poorly understood. Partial sequences of the cytochrome oxidase subunit I (COXI) gene were obtained from seven freshwater sponge species and one marine sponge. Phylogenetic analyses showed that all the freshwater sponge species formed one common clade and they appeared to be closely related. These results indicate that freshwater sponges are of monophyletic origin. Potamolepidae and Spongillidae are situated at the base of the tree and are assumed to be phylogenetically older than Lubomirskiidae.

Lake Baikal is known for its high percentage of endemic fauna and flora. The most abundant sessile animal taxa in the littoral zone of Baikal are the photosymbiotic sponges. These endemic sponges are grouped to the family Lubomirskiidae and are separated, based on molecular data, from the cosmopolitan family Spongillidae Gray 1867. In the present review, recent data on the potential driving forces of the rapid speciation in Lake Baikal have been unified. Current data suggest that the genetic repertoire of the sponges was sufficiently large to cope with the major cold events, occurring 2.8–2.5 and 1.8–1.5 Ma. It is proposed that during those periods of climatic incisions founder populations were separated from the parental cosmopolitan sponge population and developed subsequently to the array of endemic species. To clarify whether the endemic sponge fauna is indeed restricted to Lake Baikal only or whether there exist related taxa in other lakes, a collection of sponges in the 750 km distant Lake Chagytai was performed. This lake harbours unexpectedly large populations of the endemic species Baikalospongia dzhegatajensis. As the habitat of the Lake Chagytai (algal habitat) differs strongly from that of Lake Baikal (rocky habitat), it is proposed that after the formation of the initial founder population in response to the climatic shift, subsequent speciation was driven by habitat differences. Zusammenfassung Der Baikalsee ist fur seine hohe Anzahl an endemischen Tieren und Pflanzen bekannt. Die haufigsten sessilen Tiertaxa in der Littoralzone des Baikalsees sind die photosymbiontischen Schwamme. Sie werden in der Familie Lubomirskiidae zusammengefasst und auch aufgrund molekular-biologischer Analysen abgetrennt von der kosmopolitischen Familie der Spongillidae. In dem vorliegenden Ubersichtsartikel wird versucht, die Selektionsdrucke, die zu der schnellen Speziation der Schwamme im Baikalsee beigetragen haben, zu formulieren. Die vorliegenden Daten lassen annehmen, dass das genetische Repertoire der Schwamme genugend komplex war, um den Kaltperioden, die am Baikalsee beobachten wurden (vor 2,8–2,5 und 1,8–1,5 Millionen Jahren) zu widerstehen. Es wird postuliert, dass wahrend dieser einschneidenden Perioden sich Grunderpopulationen von kosmopolitischen Schwamm-Spezies separiert haben, die sich anschliesend in eine Reihe endemischer Schwamme aufgespalten haben. Zur Abklarung, ob endemische Schwamme der Region auf den Baikalsee beschrankt sind, wurden Tiere aus dem 750 km entfernten Chagytai-See untersucht. Dieser See zeichnet sich durch uberraschend grose Populationen der endemischen Schwamm-Spezies Baikalospongia dzhegatajensis aus. Das Habitat des Chagytai-Sees (Dominanz an Algen) unterscheidet sich stark vom Baikalsee. Es wird angenommen, dass nach der Ausbildung einer ursprunglichen Grunderpopulation, als Antwort auf klimatische Anderungen, Habitat-Unterschiede dazu beigetragen haben, dass sich die Speziation der Schwamme im Baikalsee und Chagytai-See unterschiedlich entwickeln konnte.

ISSN 16076729, Doklady Biochemistry an d Biophysics, 2013, Vol. 448, pp. 1–3. © Pleiades Publishing, Ltd., 2012.Original Russian Text © I.G. Kondratov, M.A. Khasnatinov, U.V. Potapova, V.V. Potapov, S.A. Levitskii, G.N. Leonova, E.V. Pavlenko, I.S. Solovarov, N.N. Denikina, N.V. Kulakova,S.I. Belikov, 2013, published in Doklady Akademii Nauk, 2013, Vol. 448, No. 1, pp. 000–000.

The phylogenetic relationship of the freshwater sponges (Porifera) in Lake Baikal is not well understood. A polyphyletic and/or monophyletic origin have been proposed. The (endemic) Baikalian sponges have been subdivided into two families: endemic Lubomirskiidae and cosmopolitan Spongillidae. In the present study, two new approaches have been made to resolve the phylogenetic relationship of Baikalian sponges; analysis of (1) nucleotide sequences from one mitochondrial gene, the cytochrome oxidase subunit I (COI) and of (2) one selected intron from the tubulin gene. Specimens from the following endemic Baikalian sponge species have been studied; Lubomirskia baicalensis, Baikalospongia intermedia, Baikalospongia recta, Baikalospongia bacillifera and Swartschewskia papyracea. They are all grouped to the family of Lubomirskiidae. Sequence comparisons were performed with the ubiquitously distributed freshwater sponge Spongilla lacustris (family Spongillidae) as well as with one marine sponge, Suberites domuncula. A sequence comparison* of the mitochondrial COI gene revealed a monophyletic grouping of the endemic Baikalian sponges with S. lacustris as the most related species to the common ancestor. The sequences of the COI gene from B. recta, B. intermedia, B. bacillifera and L. baicalensis were found to be identicaland separated from those of S. lacustris and S. papyracea. In a second approach, the exon/intron sequences framing the intron-2 of the sponge tubulin gene were chosen for the phylogenetic analysis. The intron sequences were aligned and used for construction of a phylogenetic tree. This analysis revealed again a monophyletic grouping with S. lacustris as the closest related species to the common ancestor. It is concluded that the Baikalian sponges, which have been studied here, are of monophyletic origin. Furthermore, the data suggest that the endemic species S. papyracea is the phylogenetically oldest, extant, endemic Baikalian sponge species.

In order to investigate the phylogenetic relationships among endemic Baikal sponges of the family Lubomirskiidae and to clarify their taxonomy, the sequence variation at the silicatein-encoding gene and the mtDNA intergenic region was examined. Phylogenetic analysis of the silicatein α1 gene exonic regions in six freshwater sponge species revealed considerable interspecific variability of this region. The analysis performed did not support the monophyly of the genera Lubomirskia and Baikalospongia. The mtDNA region between the COX2 and ATP6 genes was examined in five species from the Lubomirskiidae family, including multiple samples for analyzing intraspecific variations. According to the data obtained, the genus Baikalospongia was paraphyletic with respect to Lubomirskia, while B. bacilifera and B. recta did not form monophyletic groups. Molecular data indicate that taxonomy of Lubomirskiidae should be revised. It was demonstrated that, in endemic Baikal sponges, the accelerated evolution was accompanied by an increase in the length of the non-coding regions in both mitochondrial and nuclear genes.

Tick-borne encephalitis virus (TBEV) is transmitted to vertebrates by taiga or forest ticks through bites, inducing disease of variable severity. The reasons underlying these differences in the severity of the disease are unknown. In order to identify genetic factors affecting the pathogenicity of virus strains, we have sequenced and compared the complete genomes of 34 Far-Eastern subtype (FE) TBEV strains isolated from patients with different disease severity (Primorye, the Russian Far East). We analyzed the complete genomes of 11 human pathogenic strains isolated from the brains of dead patients with the encephalitic form of the disease (Efd), 4 strains from the blood of patients with the febrile form of TBE (Ffd), and 19 strains from patients with the subclinical form of TBE (Sfd). On the phylogenetic tree, pathogenic Efd strains formed two clusters containing the prototype strains, Senzhang and Sofjin, respectively. Sfd strains formed a third separate cluster, including the Oshima strain. The strains that caused the febrile form of the disease did not form a separate cluster. In the viral proteins, we found 198 positions with at least one amino acid residue substitution, of which only 17 amino acid residue substitutions were correlated with the variable pathogenicity of these strains in humans and they authentically differed between the groups. We considered the role of each amino acid substitution and assumed that the deletion of 111 amino acids in the capsid protein in combination with the amino acid substitutions R16K and S45F in the NS3 protease may affect the budding process of viral particles. These changes may be the major reason for the diminished pathogenicity of TBEV strains. We recommend Sfd strains for testing as attenuation vaccine candidates.

This study focused on finding, culturing, and identifying the biological and genetic characteristics of three louping ill virus (LIV) strains in the south of the Russian Far East. The Primorye-155-77 and Primorye-20-79 virus strains were isolated from Ixodes persulcatus ticks, and the Primorye-185-91 strain was isolated from the blood of a person after a tick bite. According to the hemagglutination and neutralization tests, Primorye-155-77, Primorye-20-79 and Primorye-185-91 had weak reactivity with antibodies in an antiserum against tick-borne encephalitis virus. In Primorye-155-77 and Primorye-20-79, the sequences of the 5' ends of the 2456-nucleotide-long viral RNA including the 5' untranslated region (UTR) and genes of the capsid protein, prM protein and envelope E protein were determined. The complete genome sequence of Primorye-185-91 was determined. The E protein gene of the Negishi strain differed from those of three analyzed strains, as there were mutations resulting in the replacement of three amino acids: Ala163Thr, Asp193Asn and Ala313Thr. The homology of Primorye-185-91 to LIV 369/T2 was 97.57 %, and to the Penrith strain, it was 98.36 %. Phylogenetic analysis showed that Primorye-155-77, Primorye-20-79 and Primorye-185-91 are related to LI/A and LI/K strains isolated in England and Scotland and to the Negishi strain; these strains have a common progenitor. Negishi-like strains were represented by one subtype of louping ill virus, i.e. the British subtype (LIV-Brit). The possibility is discussed of a single introduction of the virus to the Far Eastern region (Japan and Primorsky Krai) from a single natural locus more than 50 years ago.

We describe the biological properties and molecular characteristics of complete genomes of 33 tick-borne encephalitis virus (TBEV) strains that induced different forms of infection, from inapparent to severe focal ones resulting in fatal outcome. Hemagglutinating activity of Oshima-like strains was higher at pH 5.8, while activity of Sofjin- and Senhzang-like strains were higher at pH 6.2 and 6.8, respectively. We determined susceptibility of porcine kidney (PK) cell cultures to these TBEV strains by cytopathic effect (CPE), plaque formation, and size of plaques. The clinical TBEV strains had higher virus titers both in tissue culture infectious dose 50(TCID50) and in plaque-forming unit (PFU) titers and larger plaques than the inapparent strains. A comparison of virus multiplication kinetics by PFU in culture fluid with kinetics of ELISA antigen and hemagglutinin accumulation suggested a different mechanism of interaction between these virus strains and PK cells at the initial stage of cell infection.

A holdfast is a root- or basal plate-like structure of principal importance that anchors aquatic sessile organisms, including sponges, to hard substrates. There is to date little information about the nature and origin of sponges’ holdfasts in both marine and freshwater environments. This work, to our knowledge, demonstrates for the first time that chitin is an important structural component within holdfasts of the endemic freshwater demosponge Lubomirskia baicalensis . Using a variety of techniques (near-edge X-ray absorption fine structure, Raman, electrospray ionization mas spectrometry, Morgan–Elson assay and Calcofluor White staining), we show that chitin from the sponge holdfast is much closer to α-chitin than to β-chitin. Most of the three-dimensional fibrous skeleton of this sponge consists of spicule-containing proteinaceous spongin. Intriguingly, the chitinous holdfast is not spongin-based, and is ontogenetically the oldest part of the sponge body. Sequencing revealed the presence of four previously undescribed genes encoding chitin synthases in the L. baicalensis sponge. This discovery of chitin within freshwater sponge holdfasts highlights the novel and specific functions of this biopolymer within these ancient sessile invertebrates.

International audience; This work demonstrates that chitin is an important structural component within the skeletal fibers of the freshwater sponge Spongilla lacustris. Using a variety of analytical techniques (13 C solid state NMR, FT-IR, Raman, NEXAFS, ESI-MS, Morgan-Elson assay and Calcofluor White Staining); we show that this sponge chitin is much closer to a-chitin, known to be present in other animals, than to b-chitin. Genetic analysis confirmed the presence of chitin synthases, which are described for the first time in a sponge. The presence of chitin in both marine (demosponges and hexactinellids) and freshwater sponges indicates that this important structural biopolymer was already present in their common ancestor.

Fragments of mtDNA genes Cyt B, ATPase 6, and ATPase 8 of six cottoid fishes species of Lake Baikal (East Siberia) were amplified and sequenced. In addition mtDNAs of the same fish were subjected to restriction analysis. The data obtained were used to construct phylogenetic trees. The topology of the ATPase tree differs from those of the Res (restriction) and Cyt B trees. Clustering of species within the trees confirms the viewpoint of Taliev (1955, Baicalian Sculpins (Cottoidei)) according to which Baikalian cottoids originate from two ancestral forms. The times of branching obtained do not confirm the existing viewpoint according to which the two golomyankas (Comephorus baicalensis and Comephorus dybowskii) are pre-Baikal (Myocene) relicts: these two species may have originated 1.2-1.8 million years ago in Baikal, and they seem to represent an example of rapid morphological evolution which resulted in the formation of a new family.

Tick-borne encephalitis (TBE) is one of the most important arboviral diseases across Eurasia. TBE virus (TBEV) is transmitted by tick bite and causes a potentially fatal neurological infection in humans. In the Russian Federation, TBE is endemic in most regions, with 3000-5000 cases of the disease annually. To characterise TBEV in Eastern Siberia, the complete genomes of five TBEV isolates from patients with different clinical manifestations were sequenced. The results show that the Siberian and Far Eastern subtypes of TBEV cause the disease in people in Eastern Siberia. Complete genome analysis revealed an unexpectedly high genetic variability within the Siberian subtype.

Sponges (phylum Porifera) are phylogenetically ancient Metazoa that use silicon to form their skeletons. The process of biomineralization in sponges is one of the important problems being examined in the field of research focused on sponge biology. Primmorph cell culture is a convenient model for studying spiculogenesis. The aim of the present work was to produce a long-term primmorph culture from the freshwater Baikal sponge Lubomirskia baikalensis (class Demospongiae, order Haplosclerida and family Lubomirskiidae) in both natural Baikal water and artificial Baikal water to study the influence of silicate concentration on formation and growth of spicules in primmorphs. Silicate concentration plays an important role in formation and growth of spicules, as well as overabundance of silica leads to destruction of cell culture primmorphs. We also found that the composition of chemical elements (Si, O, C, and Na) varied along the length of growing spicules at cultivation in different media. The long-term culture of Baikal sponge primmorphs will be necessary for further investigations, and this system may serve as a powerful in vitro model to study spiculogenesis in Baikal siliceous sponges during the early stages of intracellular spicule formation to identify genes that affect biomineralization.

Two protein-coding regions (cytochrome b, ATPase 8, and part of ATPase 6) from mitochondrial DNA of Cottus kessleri, Cottocomephorus grewingki, and Cottocomephorus inermis--Baikalian endemic sculpins--were amplified via polymerase chain reaction, and sequenced. Two novel primers--L8352 (5'-TAAAGATTGGTGAC TCCCAACCACC) and H8773 (5'-GTAGGGAGT AAGCCCAATATGTT)--were used for the latter region. Phylogenies suggested by sequence divergence of the genes of ATPases appeared to be different from those computed from data for cytochrome b. The time of species branching was estimated as 1-2 million years (Myr) on the basis of merged sequences. Hence, members of the Baikalian cottoid species flock are much more distant from each other than members of the cichlid fish flocks of the great lakes of Africa (0.2 Myr). Topology of the phylogenetic tree does not contradict the relationships derived from morphological data. However, genetic distances suggest that C. grewingki and C. inermis are not sister species, contrary to general belief.