Diversity of the major capsid genes (g23) of T4-like bacteriophages in the eutrophic Lake Kotokel in East Siberia, Russia

Numerous studies revealed high diversity of T4-like bacteriophages in various environments, but so far, little is known about T4-like virus diversity in freshwater bodies, particularly in eutrophic lakes. The present study was aimed at elucidating molecular diversity of T4-like bacteriophages in eutrophic Lake Kotokel located near Lake Baikal by partial sequencing of the major capsid genes (g23) of T4-like bacteriophages. The majority of g23 fragments from Lake Kotokel were most similar to those from freshwater lakes and paddy fields. Despite the proximity and direct water connection between Lake Kotokel and Lake Baikal, g23 sequence assemblages from two lakes were different. UniFrac analysis showed that uncultured T4-like viruses from Lake Kotokel tended to cluster with those from the distant lake of the same trophic status. This fact suggested that the trophic conditions affected the formation of viral populations, particularly of T4-like viruses, in freshwater environments.     

Presence and genetic diversity of microcystin-producing cyanobacteria (<Emphasis Type="Italic">Anabaena</Emphasis> and <Emphasis Type="Italic">Microcystis</Emphasis>) in Lake Kotokel (Russia,Lake Baikal Region)

A survey was conducted for the presence of cyanobacteria toxins in Lake Kotokel due to a few cases of Haff disease registered in 2008–2009 caused by consumption of fish from Lake Kotokel, and wildlife mortality including large fish kill. The aims of this study were to determine what cyanotoxins (if any) were present in the lake, to describe phytoplankton composition including morphology, density, and species diversity of cyanobacteria, as well as to evaluate the trophic state of the lake. Samples were collected from both nearshore and central sites in August of 2009. Aphanocapsa holsatica dominated the phytoplankton. The presence of toxigenic genotypes of Microcystis spp. and Anabaena lemmermannii was detected by sequencing of PCR-amplified aminotransferase domain of microcystin synthetase gene. LR, RR, and YR microcystin (MC) variants were detected with liquid chromatography-UV mass spectrometry. The data do not shed light on the etiology of Haff disease in Lake Kotokel region, nevertheless taking into account the recreational importance of the lake and its direct connection to Lake Baikal, a necessity to monitor cyanobacteria in these water bodies is evident. This is the first report on simultaneous detection of MC-producing genotypes and MCs in the Lake Baikal region.     

Ultrastructure of phagocytic cells in the spleen of Psammophis sibilans (serpentes: Colubridae)

The spleen of Psammophis sibilans is composed mainly of red pulp, the white pulp being poorly developed. The white pulp lymphoid clusters are scattered throughout the organ and contain lymphocytes, reticular cells, and some plasma cells. The red pulp consists of reticular cells intermingled with blood cells, sinusoids, and melanomacrophage centers (MMCs). Filtering of particulate matter from the blood occurs in the red pulp by phagocytes of the pulp cord. MMCs are formed by the association of free macrophages that have phagocytosed some blood cells. Early filtering of particulate matter by the phagocytes of the pulp cords may allow for more efficient phagocytosis of erythrocytes by the MMCs. © 1994 Wiley-Liss, Inc.     

Functional dissimilarity of melanomacrophage centres in the liver and spleen from females of the teleost fish <Emphasis Type="Italic">Prochilodus argenteus</Emphasis>

Melanomacrophage centres (MMCs) are formed by macrophage aggregates containing pigments such as hemosiderin, melanin and lipofuscin. MMCs are found in animals such as reptiles, amphibians and, mainly, fishes, in organs such as the kidney, spleen, thymus and liver. In teleost fish, several functions have been attributed to MMCs, including the capture and storage of cations, the phagocytosis of cellular debris and immunological reactions. As the use of MMCs has been suggested as a tool for the assessment of environmental impacts, our aim has been to describe the various metabolic processes performed by MMCs in diverse organs (liver and spleen) by using the teleost Prochilodus argenteus as an animal model. MMCs from the liver and spleen were assessed by histochemistry, transmission electron microscopy, scanning electron microscopy, X-ray microanalysis techniques and biochemical assay for N-acetylglucosaminidase activity. The data showed metabolic differences in MMCs between the liver and spleen of P. argenteus in their morphometric characteristics and biochemical and elemental composition. The implications of these findings are discussed, focusing on their role in organ metabolism.     

Structure of the spleen of the sea bass (Dicentrarchus labrax): A light and electron microscopic study

The spleen of sea bass (Dicentrarchus labrax) is composed mainly of red pulp, whereas the white pulp is poorly developed. The red pulp consists of clear reticular cells intermingled with blood cells, sinusoids, and melanomacrophage centers (MMCs). The MMCs are enclosed by an interrupted connective tissue capsule and show some areas in continuity with the adjacent pulp. The MMCs are formed by the association of free macrophages that have phagocytosed some blood cells. Sparse white pulp is diffuse, forming a cuff around the pulp arteries and MMCs, or occurring in small groups between the splenic cords. A longitudinal artery and vein, lying side by side, extend the length of the spleen. Frequently the capillaries are surrounded by a sheath of macrophages or ellipsoids. These macrophages may contain erythrocytes in varying degrees of degradation. Lymphopoiesis and plasmapoiesis occur in the sparse lymphold areas. Abundant plasma cell groups may indicate the presence of antibody production.     

Karyotypic features of Chironomus entis and Chironomus borokensis (Diptera, Chironomidae) from Lake Kotokel (Lake Baikal Basin)

Karyotypes and chromosomal polymorphism of sympatric sibling species Chironomus entis and Chironomus borokensis from Lake Kotokel were studied. Five inversion banding sequences were found in C. entis, including p’entA16, p’entB9, and p’entF6 that are new for the species; in C. borokensis, four inversion banding sequences were revealed, borA4 being new for the species. The level of chromosome polymorphism in C. entis and C. borokensis can be considered similar: 54.2 and 53.8% heterozygosity, 0.67 and 0.73 inversions per individual, respectively. The new data are compared with those on C. entis population from Lake Kotokel obtained in 1981 and those of C. borokensis from Lake Dukhovoe (Lake Baikal basin) obtained in 1983.     

The spleen of the African lungfish Protopterus annectens: freshwater and aestivation

We describe the structure of the spleen of the African lungfish Protopterus annectens in freshwater conditions, and after 6?months of aestivation. The spleen is formed by cortical tissue that surrounds the splenic parenchyma. The cortex is a reticulum that contains two types of granulocytes, developing and mature plasma cells, and melanomacrophage centres (MMCs). The parenchyma is divided into lobules that show a subcapsular sinus and areas of red pulp and white pulp. Red pulp contains vascular sinuses and atypical cords formed by delicate trabeculae. White pulp also contains vascular sinuses and cords. Structural data indicate that red pulp is involved in erythropoiesis, destruction of effete erythrocytes, and plasma cell differentiation. White pulp appears to be involved in the production of immune responses. Macrophages and sinus endothelial cells constitute the reticulo-endothelial system of the spleen. After aestivation, the number of MMCs increases, and spleen tissue is infiltrated by lymphocytes, granulocytes, and monocytes. Also, white pulp is reduced, and sinus endothelial cells undergo vacuolar degeneration. Lungfish spleen shares structural characteristics with secondary lymphoid organs of both ectothermic and endothermic vertebrates, but appears to have evolved in unique ways.     

Immunocytochemical detection of Ig-positive cells in blood, lymphoid organs and the gut associated lymphoid tissue of the turbot (Scophthalmus maximus)

The present study was designed to search for the sites of the B-cell lineage in the different lymphoid organs of turbot (Scophthalmus maximus) by immunoperoxidase staining with a rabbit polyclonal antiserum against deglycosylated turbot IgM (TUDG-6). A turbot immunoglobulin (Ig) fraction, isolated by protein A, was checked for purity by gel filtration and SDS-PAGE under reducing conditions. The turbot IgM was deglycosylated and used to raise an antiserum. The antiserum titre was evaluated in ELISA. It was then used to analyse turbot peripheral blood leucocytes for membrane and cytoplasmic Ig and for immunohistochemistry with turbot lymphoid tissues. Very low numbers of Ig+ cells were found in thymus sections. In sections of spleen, Ig+ cells were observed in white pulp, around ellipsoids but were mostly concentrated and associated with melanomacrophage centers (MMCs). The lymphoid Ig+ cells in the kidney tended to be dispersed among haematopoietic and granulopoietic cell populations and were in intimate association with the MMCs and blood vessels. This association between MMCs and Ig+ cells in the spleen and the kidney, is discussed with respect to the role played by these organs in the immune system of fish. Last, the lymphoid population in the gut associated lymphoid tissue (GALT) of turbot was characterised with respect to staining for Ig. Immunoreactive cells were rarely detected in the epithelial layer although many lymphocytes were present, but they were frequently observed in the lamina propria, presumably as part of the GALT and involved in mucosal immune responses.     

A model for acute iron overload in sea bass (Dicentrarchus labrax L.)

Evaluation of several parameters involved in iron metabolism was carried out after intraperitoneal (i.p.) injection with iron dextran (IDx) in sea bass (Dicentrarchus labrax L.). After treatment, a rapid mobilization of IDx from the peritoneal cavity to other organs was observed. This was followed by a modification of normal peripheral blood iron parameters. Total iron (TI) and transferrin saturation (TS) rose rapidly, to 4.14 microg/ml and 83.7%, respectively, on day 3. In contrast, unsaturated iron binding capacity (UIBC) dropped from 3.19 microg/ml (at day 0) to 0.90 microg/ml on day 3. Tissue iron content was determined by atomic absorption spectometry (AAS). Three days post-IDx injection, values of iron concentration in liver, spleen and head kidney were significantly higher than control values (15, 6 and 9-fold increase, respectively). Samples of liver, spleen and head kidney were processed for routine histology, and the Perl's method was used for iron staining. Histological sections of the IDx-treated animals showed iron deposition in all tissues studied. In the liver, the iron was evenly distributed over the whole organ, being present in the hepatocytes. In the head kidney and spleen, the iron deposition was mainly observed in the melanomacrophage centres (MMCs). The present study characterizes several parameters involved in iron metabolism, and develops a fish model, of iron overload, which can be used in further studies of iron toxicity and iron-induced susceptibility to bacterial infections.     

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

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

Heavy metal content and response of the blood system of Siberian dace <Emphasis Type="Italic">Leuciscus leuciscus baicalensis</Emphasis> (Cypriniformes,Cyprinidae) under anthropogenic impact

Chemical, hematological, and immunological characteristics of the Siberian dace Leuciscus baicalensis from different areas of Lake Baikal basin were first determined. It was established that the most considerable changes in hemopoesis, cellular, and humoral links of immunity were in individuals with a high content of heavy metals in the liver from populations inhabiting the Selenga River downstream the Ulan Ude Industrial Center and the Selenga Pulp-and-Cardboard Plant. Changes in immunohematological responses of L. leusiscus baicalensis from Cherkalov Bay of Lake Baikal proceeded within the adaptive possibilities of the species and indicated the development of compensatory reactions in response to the accumulation of heavy metals in the body. The results obtained make it possible to assert that Chivurkuiskii Bay of Baikal may serve as a control polygon during monitoring of immunobiological states of fish populations from Baikal basin.     

Phytomonitoring of air pollution in the Baikal region

Phytomonitoring of air pollution has been performed using the Scots pine as a highly sensitive bioindicator. Needles have been sampled from key plots along the macrotransect Sayansk-Irkutsk-Listvyanka (with a total length of 320 km) covering the main industrial centers of the Baikal region and stretching up to the shoreline of Lake Baikal. The content of inorganic pollutants (sulfur, fluorides, heavy metals, etc.) has been determined. The highest level of air pollution has been registered in the following industrial centers: Angarsk, Irkutsk, Usol’e-Sibirskoe, and Shelekhov. The degree of pollution is lower in industrial centers such as Sayansk and Cheremkhovo. The data provide evidence that most pollutants reach the shoreline of Lake Baikal.     

Melanomacrophage Aggregations and Their Age Relationships in the Yellow Mud Turtle,Kinosternon flavescens (Kinosternidae)

Microscopic examination of the pigment cell aggregations in the liver and spleen of mud turtles (Kinosternon flavescens) suggests that the morphology and behavior of these cells is consistent with the melanomacrophages described in teleost fishes and the pigmented “Kupffer cells” described in frogs and reptiles. These cells contain massive amounts of melanin, substantial lipofuscin, and some hemosiderin consistent with their phagocytic function. Similar-appearing isolated pigmented macrophages are solitary in the liver, spleen, lung, and kidney. Number and size of the largest hepatic aggregations increase almost linearly with turtle age so that in old turtles they may constitute up to 20% of the liver volume. This increase may result from hepatic recruitment of macrophages throughout the life of the turtle and suggests that size and number of melanomacrophage aggregations may serve as a marker for senescence in otherwise healthy turtles of this species.     

Mercury content in muscles of fish of the Selenga River and lakes of its basin (Russia)

Differences between the muscle mercury contents in fish from lakes Gusinoye and Karasinoye and the Selenga River may indicate an inconsiderable income of mercury in aquatic ecosystems with atmospheric precipitation, along with a more intensive migration of mercury with the riverine flux. The minimal concentration of mercury (less than 0.15 mg/kg, dry weight) has been registered in the muscles of Amur sleeper from Lake Karasinoye and of Baikal omul from Lake Baikal, while the maximal concentration (1.0–2.3 mg/kg, dry weight) has been revealed in the fish caught in the Selenga River delta (in predatory pike, Amur catfish, and perch, as well as in omnivorous roach and ide). The muscle metal content in dace decreased along the downstream direction.     

Lympho-granulocytic tissue associated with the wall of the spiral valve in the African lungfish Protopterus annectens

We describe the structure of the lympho-granulocytic tissue associated with the wall of the spiral valve of the African lungfish Protopterus annectens. The study was performed under freshwater conditions and after 6 months of aestivation. The lympho-granulocytic tissue consists of nodes surrounded by reticular tissue. The nodes are formed by an outer and an inner component separated by a thin collagenous layer. The outer component is a reticular-like tissue that contains two types of granulocytes, developing and mature plasma cells and melanomacrophage centres (MMCs). The inner component, the parenchyma, contains a meshwork of trabeculae and vascular sinusoids and shows dark and pale areas. The dark areas contain diffuse lymphoid tissue, with a large number of mitoses and plasma cell clusters. The pale areas contain a small number of macrophages and lymphocytes. Macrophages and sinus endothelial cells are filled with haemosiderin granules and appear to form part of the reticuloendothelial system of the lungfish. The reticular tissue houses granulocytes, plasma cells and MMCs and might serve for the housing and maturation of cells of the white series. After aestivation, the nodes undergo lymphocyte depletion, the suppression of mitosis, granulocyte invasion and the occurrence of cell death. By contrast, few histological changes occur in the reticular tissue. Whereas the nodes appear to be involved in lymphocyte proliferation and plasma cell maturation, the function of the reticular tissue remains obscure.     

Dependence that the community structure of the roach <Emphasis Type="Italic">Rutilus rutilus</Emphasis> (L.) parasites has on the host age

The changes that take place in the structure of the parasite communities of two roach populations as the hosts grow older have been studied in the Chivyrkuiskii Bay of Lake Baikal and the estuary of the Selenga River. The parasite communities are analyzed at the levels of host individuals (infracommunities), host age groups (sets of infracommunities), and host population (component community). The number of parasite species regularly increases as the roaches age in both aquatic bodies, whereas the evenness of the parasite communities decreases. The Berger-Parker dominance index of the parasite communities changes with the roach age in different directions, increasing in the Chivyrkuiskii Bay and decreasing in the estuary of the Selenga River. The changes in the Shannon diversity index for sets of infracommunities of roach parasites are the opposite of those in the Berger-Parker index.     

Diatom succession trends in recent sediments from Lake Baikal and their relation to atmospheric pollution and to climate change

Recent environmental change in Lake Baikal has been attributed to anthropogenic influences on the ecosystem, especially through pollution and cultural eutrophication. These hypotheses are tested in this paper principally by diatom analyses in 20 short sediment cores. Most of the cores were collected with a new type of box corer specifically designed for use on Lake Baikal. Most cores contain a good sediment record but turbidites occur in some sediment profiles which may be best recognized using a combination of techniques, such as radiometric dating and percentage dry weight analyses. The most recent sediments, especially those in the southern basin and in the very north of Baikal, contain a record of anthropogenic contamination in the form of lead and spheroidal carbonaceous particles, which confirms that the southern basin of Baikal is most affected by atmospheric sources of pollution. However, there is no sedimentary diatom evidence indicating offshore water quality deterioration in Baikal owing to air pollution or eutrophication. Small increases in diatoms which indicate nutrient enrichment (e.g. Stephanodiscus minutulus, Synedra acus v. radians and Synedra acus v. acus) may reflect local eutrophication of the shallow waters close to the Selenga Delta and certain coastal sites in the southern basin near to the Baikalsk paper and pulp mill. By using numerical techniques, Lake Baikal can be split into at least four regions on the basis of its surface sediment flora: the south, middle and north basins, and the shallow waters surrounding the Selenga Delta region. Diatom analyses reveal that the endemic flora of Lake Baikal has been constantly changing over at least the last 2000 years and that these fluctuations are probably responses to natural climatic variability. Recent sediments of Baikal may be affected by taphanomic processes (e.g. dissolution) and turbidite deposition, and these must be taken into account when interpreting the sedimentary diatom record. The diatom flora of the lake is currently dominated by several species, such as Aulacoseira baicalensis, A. islandica, Cyclotella minuta and Stephanodiscus binderanus v. baicalensis. All these species, except for C. minuta, have become more common in the lake in approximately the last 130 years, and we hypothesize that these changes may be attributed to a number of different processes linked to an ameliorating climate after the end of the Little Ice Age. The results presented here have important implications for this recently designated World Heritage Site, with regard to future pollution controls and catchment management policies.     

Centric diatoms from Lake Frolikha (Transbaikal area) and peculiarities of distribution of some taxa in Asia

The species composition of centric diatoms from Lake Frolikha (Transbaikal area) has been studied. The lake is located on the eastern shore of Lake Baikal and connected with it by a river. Twenty-three species of Centrophyceae from 7 genera (Aulacoseira, Cyclostephanos, Cyclotella, Discostella, Handmania, Pliocaenicus, and Stephanodiscus) have been found. The most represented genus is Aulacoseira (11 species). Fifteen species and 4 genera are new for the lake record. All revealed species are known in other lakes of the Baikal region; however Baikal endemics are absent in Lake Frolikha. The flora of centric diatoms in Lake Frolikha can be divided into two groups. The first group includes taxa common in Lake Baikal, and the second group includes taxa not typical for Lake Baikal. The level of differences between flora in Lake Baikal and Lake Frolikha is high (43%) despite the close location and connection to the river. An analysis of distribution of interesting species in Asia is presented.     

Spectral tuning and evolution of short wave-sensitive cone pigments in cottoid fish from Lake Baikal

The cottoid fishes of Lake Baikal in eastern Siberia provide a unique opportunity to study the evolution of visual pigments in a group of closely related species exposed to different photic environments. Members of this species flock are adapted to different depth habitats down to >1000 m, and both the rod and cone visual pigments display short wave shifts as depth increases. The blue-sensitive cone pigments of the SWS2 class cluster into two species groups with lambda(max) values of 450 and 430 nm, with the pigment in Cottus gobio, a cottoid fish native to Britain, forming a third group with a lambda(max) of 467 nm. The sequences of the SWS2 opsin gene from C. gobio and from two representatives of the 450 and 430 nm Baikal groups are presented. Approximately 6 nm of the spectral difference between C. gobio and the 450 nm Baikal group can be ascribed to the presence of a porphyropsin/rhodopin mixture in C. gobio. Subsequent analysis of amino acid substitutions by site-directed mutagenesis demonstrates that the remainder of the shift from 461 to 450 nm arises from a Thr269Ala substitution and the shift from 450 to 430 nm at least partly from Thr118Ala and Thr118Gly substitutions. The underlying adaptive significance of these substitutions in terms of spectral tuning and signal-to-noise ratio is discussed.     

Colorless sulfur bacteria <Emphasis Type="Italic">Thioploca</Emphasis> from different sites in Lake Baikal

The colorless sulfur bacteria Thioploca spp. found in Lake Baikal are probably a marker for the influx of subterranean mineralized fluids. Bacteria act as a biological filter; by consuming sulfide in their metabolism, they detoxicate it and maintain the purity of Lake Baikal’s water. The bacteria were investigated by various techniques. According to analysis of the 16S rRNA gene fragment, Thioploca sp. from Frolikha Bay, Baikal belongs to the clade of freshwater species found in Lake Biwa and Lake Constance; it is most closely related to Thioploca ingrica.