The low mitochondrial diversities in lizardfish Saurida elongate: Recent population expansion and selection

To investigate the phylogeographic pattern and historical demography of Saurida elongata, a 463 base pair fragment of the mitochondrial DNA control region was analyzed in 118 individuals collected from five populations in the East China Sea and South China Sea. The results revealed remarkably low nucleotide diversity in S. elongata, which was probably caused by selection during Pleistocene drastic climate fluctuations. Phylogenetic relationship was continuous, but a localized clade was found dominating Beihai population. Mismatch analyses showed that S. elongata experienced both demographic and range expansions. Among these populations, Beihai population expanded latest. Localization of haplotypes clade in Beihai was likely a result of the selection when new habitat was established during a range expansion, and indicated limited gene flow between Beihai and other populations. Our results suggested Beihai population should be managed separately in fishery.     

Coexistence of Bacterial Sulfide Oxidizers, Sulfate Reducers, and Spirochetes in a Gutless Worm (Oligochaeta) from the Peru Margin

Olavius crassitunicatus is a small symbiont-bearing worm that occurs at high abundance in oxygen-deficient sediments in the East Pacific Ocean. Using comparative 16S rRNA sequence analysis and fluorescence in situ hybridization, we examined the diversity and phylogeny of bacterial symbionts in two geographically distant O. crassitunicatus populations (separated by 385 km) on the Peru margin (water depth, ~300 m). Five distinct bacterial phylotypes co-occurred in all specimens from both sites: two members of the γ-Proteobacteria (Gamma 1 and 2 symbionts), two members of the δ-Proteobacteria (Delta 1 and 2 symbionts), and one spirochete. A sixth phylotype belonging to the δ-Proteobacteria (Delta 3 symbiont) was found in only one of the two host populations. Three of the O. crassitunicatus bacterial phylotypes are closely related to symbionts of other gutless oligochaete species; the Gamma 1 phylotype is closely related to sulfide-oxidizing symbionts of Olavius algarvensis, Olavius loisae, and Inanidrilus leukodermatus, the Delta 1 phylotype is closely related to sulfate-reducing symbionts of O. algarvensis, and the spirochete is closely related to spirochetal symbionts of O. loisae. In contrast, the Gamma 2 phylotype and the Delta 2 and 3 phylotypes belong to novel lineages that are not related to other bacterial symbionts. Such a phylogenetically diverse yet highly specific and stable association in which multiple bacterial phylotypes coexist within a single host has not been described previously for marine invertebrates.     

The diversity, antimicrobial and anticancer activity of endophytic fungi associated with the medicinal plant Stryphnodendron adstringens (Mart.) Coville (Fabaceae) from the Brazilian savannah

The diversity and biological activities of the endophytic fungi associated with the Brazilian medicinal plant Stryphnodendron adstringens were studied. A total of 320 fungal isolates were obtained, and 66 phylotypes comprising 25 genera were identified. The fungal community of S. adstringens displayed high richness, diversity and low dominance indices. The most abundant phylotypes were closely related to Diaporthe phaseolorum, Guignardia camelliae, and Preussia pseudominima. Sixteen fungal extracts displayed biological activities when screened against bacteria, fungi, cancer cell lines, and amastigote forms of Leishmania amazonensis. The extract of phylotype Nigrospora cf. oryzae exhibited a selective antifungal activity and inhibited the growth of Candida albicans and Cladosporium sphaerospermum. The extracts of Diaporthe cf. phaseolorum and Xylaria sp. phylotypes displayed anticancer activities. Our results indicate that the endophytes associate with this medicinal plant may be a source for novel drugs.     

Niche Partition of Bacteriovorax Operational Taxonomic Units Along Salinity and Temporal Gradients in the Chesapeake Bay Reveals Distinct Estuarine Strains

The predatory Bacteriovorax are Gram-negative bacteria ubiquitous in saltwater systems that prey upon other Gram-negative bacteria in a similar manner to the related genus Bdellovibrio. Among the phylogenetically defined clusters of Bacteriovorax, cluster V has only been isolated from estuaries suggesting that it may be a distinct estuarine phylotype. To assess this hypothesis, the spatial and temporal distribution of cluster V and other Bacteriovorax phylogenetic assemblages along the salinity gradient of Chesapeake Bay were determined. Cluster V was expected to be found in significantly greater numbers in low to moderate salinity waters compared to high salinity areas. The analyses of water and sediment samples from sites in the bay revealed cluster V to be present at the lower salinity and not high salinity sites, consistent with it being an estuarine phylotype. Cluster IV had a similar distribution pattern and may also be specifically adapted to estuaries. While the distribution of clusters V and IV were similar for salinity, they were distinct on temperature gradients, being found in cooler and in warmer temperatures, respectively. The differentiation of phylotype populations along the salinity and temporal gradients in Chesapeake Bay revealed distinct niches inhabited by different phylotypes of Bacteriovorax and unique estuarine phylotypes.     

Results of testing the comparatory method: The curvature of the shell valve frontal section is inappropriate as a systematic character for the freshwater pearl mussel of the genus Margaritifera

This paper continues a discussion on the number of pearl mussel species of the genus Margaritifera in northern Europe. A biometric study of 1711 pearl mussel Margaritifera margaritifera shells from 15 rivers in Russia and Latvia (basins of the White and Baltic seas) has been conducted. All the examined samples fall into two groups: the northern group (with the shells more flattened on average, f. margaritifera) and the southern one (with more convex shells, f. elongata); the boundary between these groups is at 63° N. Analysis of intrapopulation variation has shown that the samples contain individuals that correspond to f. margaritifera, f. elongata, and f. borealis. However, any hiatus between these forms is absent in all the samples, and individuals belonging to two intermediate forms are rather frequent. The hypothesis on the species specificity of the shell valve frontal section has not been confirmed based on examination of large shell samples. The pearl mussels inhabiting rivers of Northern Europe belong to a single species, M. margaritifera.     

Dominant phylotypes in the 16S rRNA gene clone libraries from bacterial mats of the Uzon caldera (Kamchatka,Russia) hydrothermal springs

In situ analysis of the 16S rRNA genes from bacterial mats of five hydrothermal springs (36–58°C) in the Uzon caldera (Kamchatka, Russia) was carried out using clone libraries. Eight clone libraries contained 18 dominant phylotypes (over 4–5%). In most clone libraries, the phylotype of the green sulfur bacterium Chlorobaculum sp. was among the dominant ones. The phylotypes of the green nonsulfur bacteria Chloroflexus and Roseiflexus and of purple nonsulfur bacteria Rhodoblastus, Rhodopseudomonas, and Rhodoferax were also among the dominant ones. Cyanobacteria were represented by one dominant phylotype in a single spring. Among nonphototrophic bacteria, the dominant phylotypes belonged to Sulfyrihydrogenibium sp., Geothrix sp., Acidobacterium sp., Meiothermus sp., Thiomonas sp., Thiofaba sp., and Spirochaeta sp. Three phylotypes were not identified at the genus level. Most genera of phototrophic and nonphototrophic organisms corresponding to the phylotypes from Uzon hydrotherms have been previously revealed in the hydrotherms of volcanically active regions of America, Asia, and Europe. These results indicate predominance of bacterial mats carrying out anaerobic photosynthesis in the hydrotherms of the Uzon caldera.     

Eukaryotic phylotypes in aquatic moss pillars inhabiting a freshwater lake in East Antarctica, based on 18S rRNA gene analysis

Aquatic mosses of Leptobryum species form unique tower-like pillars of vegetation termed “moss pillars” in Antarctic lakes. Moss pillars have distinct redox-affected sections: oxidative exteriors and reductive interiors. We have proposed that a “pillar” is a community and habitat of functionally interdependent organisms and may represent a mini-biosphere. Batteries of 16S rRNA genotypes, or phylotypes, of eubacteria and cyanobacteria, but no archaea, have been identified in moss pillars. However, detailed identification or phylogenetic analyses of the moss and their associated eukaryotic microbiota have not been performed. This study analyzed near-full-length 18S rRNA gene sequences obtained from two whole moss pillars. In total, 28 PCR clone libraries from two whole moss pillars were constructed, and 96 clones from each library (total 2,688 clones) were randomly selected and sequenced. Molecular phylogenetic analysis revealed that the phylotype belonging to Bryophyta, considered to be derived from moss, was closely related (99.9?%) to the 18S rRNA gene sequence from Leptobryum pyriforme. Unexpectedly, phylotypes belonging to a novel clade of fungi dominated (approximately 27–75?%) the moss pillar libraries. This suggests that fungi may contribute to carbon cycling in the moss pillar as parasites or decomposers. In addition, phylotypes related to ciliates and tardigrades were subdominant in the exterior, while the phylotype of the ameba-like, single-celled eukaryote, Cercomonas (Cercozoa), was detected only in the interior. These features were shared by both moss pillars. The 18S rRNA gene-based profiles demonstrated that redox-related factors may control distribution of some eukaryotic microbes in a whole moss pillar.     

Identification of Desulfosporosinus as toluene-assimilating microorganisms from a methanogenic consortium

To understand the potential for toluene removal under electron acceptor depleted conditions, stable isotope probing (SIP) was applied to a methanogenic toluene degrading culture to identify the microorganisms responsible for toluene assimilation. Both bacterial and archaeal communities were investigated. The approach involved addition of labeled and unlabeled toluene to microcosms, DNA extraction, ultracentrifugation, and analysis of the generated fractions, as well as the total genomic DNA. Three genes were investigated in the fractions, including the 16S rRNA gene, bssA (encoding for benzylsuccinate synthase α-subunit) and bamA (encoding for 6-oxocylcohex-1-ene-1-carbonyl-CoA hydrolase). Analysis of the total genomic 16S rRNA gene clone library indicated the microbial community was reasonably diverse, containing microorganisms from six phyla (Proteobacteria, Firmicutes, Acidobacteria, Actinobacteria, Deferribacteres, Bacteroidetes). In contrast, only four phylotypes were found in the heavy fraction 16S rRNA gene clone library (from three phyla: Firmicutes, Acidobacteria, Actinobacteria). When these data were correlated with the TRFLP fragments enriched in the heavy fractions, three phylotypes were identified. Specifically, a Desulfosporosinus phylotype was highly enriched in the heavy fractions and was therefore the key consumer of the labeled carbon from toluene. Two other phylotypes, Peptostreptococcaceae and Pseudonocardia were presumed to consume daughter products and produce methane precursors, which in turn were likely utilized by Methanomicrobia to produce methane. Further, the SIP results suggested that the enzymes encoding by functional genes (bssA and bamA) were likely to be harbored by the Desulfosporosinus phylotype.     

Community Dynamics of Arbuscular Mycorrhizal Fungi in High-Input and Intensively Irrigated Rice Cultivation Systems

Application of a mycorrhizal inoculum could be one way to increase the yield of rice plants and reduce the application of fertilizer. We therefore studied arbuscular mycorrhizal fungi (AMF) in the roots of wetland rice (Oryza sativa L.) collected at the seedling, tillering, heading, and ripening stages in four paddy wetlands that had been under a high-input and intensively irrigated rice cultivation system for more than 20 years. It was found that AMF colonization was mainly established in the heading and ripening stages. The AMF community structure was characterized in rhizosphere soils and roots from two of the studied paddy wetlands. A fragment covering the partial small subunit (SSU), the whole internal transcribed spacer (ITS), and the partial large subunit (LSU) rRNA operon regions of AMF was amplified, cloned, and sequenced from roots and soils. A total of 639 AMF sequences were obtained, and these were finally assigned to 16 phylotypes based on a phylogenetic analysis, including 12 phylotypes from Glomeraceae, one phylotype from Claroideoglomeraceae, two phylotypes from Paraglomeraceae, and one unidentified phylotype. The AMF phylotype compositions in the soils were similar between the two surveyed sites, but there was a clear discrepancy between the communities obtained from root and soil. The relatively high number of AMF phylotypes at the surveyed sites suggests that the conditions are suitable for some species of AMF and that they may have an important function in conventional rice cultivation systems. The species richness of root-colonizing AMF increased with the growth of rice, and future studies should consider the developmental stages of this crop in the exploration of AMF function in paddy wetlands.     

Arbuscular mycorrhizal and dark septate endophytic fungi at 5,500 m on a glacier forefront in the Qinghai-Tibet Plateau, China

Arbuscular mycorrhizal (AM) and dark septate endophytic (DSE) fungi are rarely studied at extremely high elevations. Here, AM and DSE colonization in two dominant plant species (Melandrium apetalum and Poa litwinowiana) were microscopically observed on the forefront of Zhadang Glacier (5,500 m above sea level) in the Qinghai-Tibet Plateau, China. The AM fungal taxa were also identified by molecular methods. Both AM and DSE fungi synchronously colonized these two plant species, but AM dominated in M. apetalum and DSE dominated in P. litwinowiana. A total of five AM fungal spore morphotypes (Acaulospora capsicula, Diversispora sp., Glomus constrictum, G. eburneum and Glomus sp.) were found in the rhizosphere soils. Molecular identification revealed two AM fungal phylotypes: one Claroideoglomus phylotype from roots and one Diversispora phylotype from spores. These results extend the elevation at which both AM and DSE are known to occur.     

Communities of arbuscular mycorrhizal fungi in Stipa krylovii (Poaceae) in the Mongolian steppe

We examined arbuscular mycorrhizal (AM) fungi colonizing the roots of Stipa krylovii, a grass species dominating the grasslands of the steppe zone in Hustai and Uvurkhangai in Mongolia. The AM fungal communities of the collected S. krylovii roots were examined by molecular analysis based on the partial sequences of a small subunit of ribosomal RNA gene as well as AM fungal colonization rates. Almost all AM fungi detected were in Glomus-group A, and were divided into 10 phylotypes. Among them, one phylotype forming a clade with G. intraradices and G. irregulare was the most dominant. Furthermore, it was also found that most of the phylotypes include AM fungi previously detected in high altitude regions in the Eurasian Continent. Significant correlations were found among soil total N, total plant biomass and AM fungal colonization ratio, which suggested that higher plant biomass may be required for the proliferation of AM fungi in the environment. Meanwhile, redundancy analysis on AM fungal distribution and environmental variables suggested that the effect of plant biomass and most soil chemical properties on the AM fungal communities were not significant.     

Significant difference in mycorrhizal specificity between an autotrophic and its sister mycoheterotrophic plant species of Petrosaviaceae

Petrosaviaceae is a monocotyledonous plant family that comprises two genera: the autotrophic Japonolirion and the mycoheterotrophic Petrosavia. Accordingly, this plant family provides an excellent system to examine specificity differences in mycobionts between autotrophic and closely related mycoheterotrophic plant species. We investigated mycobionts of Japonolirion osense, the sole species of the monotypic genus, from all known habitats of this species by molecular identification and detected 22 arbuscular mycorrhizal (AM) fungal phylotypes in Archaesporales, Diversisporales, and Glomerales. In contrast, only one AM fungal phylotype in Glomerales was predominantly detected from the mycoheterotrophic Petrosavia sakuraii in a previous study. The high mycobiont diversity in J. osense and in an outgroup plant, Miscanthus sinensis (Poaceae), indicates that fungal specificity increased during the evolution of mycohetrotrophy in Petrosaviaceae. Furthermore, some AM fungal sequences of J. osense showed >99 % sequence similarity to the dominant fungal phylotype of P. sakuraii, and one of them was nested within a clade of P. sakuraii mycobionts. These results indicate that fungal partners are not necessarily shifted, but rather selected for in the course of the evolution of mycoheterotrophy. We also confirmed the Paris-type mycorrhiza in J. osense.     

A new species of the genus Paralomis (Crustacea: Decapoda: Lithodidae) from the Spiess seamount near Bouvet Island (Southern Ocean), with notes on habitat and ecology

A new species of the genus Paralomis, Paralomis elongata, has been collected from the Spiess seamount near Bouvet Island in the Southern Ocean. The species shows close affinity with P. anamerae Macpherson, 1988, from the Patagonian Shelf, P. africana Macpherson, 1987, from the south-western African shelf, and P. aculeata Henderson, 1888, from Crozet Islands. Morphological differences among the species and ecological characteristics are discussed.     

Phylogenetic characterization of endosymbionts of the hydrothermal vent mussel Bathymodiolus azoricus by analysis of the 16S rRNA, cbbL, and pmoA genes

In order to assess the phylogenetic diversity of the endosymbiotic microbial community of the gills of marine bivalve Bathymodiolus azoricus, total DNA was extracted from the gills. The PCR fragments corresponding to the genes encoding 16S rRNA, ribulose-bisphosphate carboxylase (cbbL), and particulate methane monooxygenase (pmoA) were amplified, cloned, and sequenced. For the 16S rDNA genes, only one phylotype was revealed; it belonged to the cluster of thiotrophic mytilid’s symbionts within the Gammaproteobacteria. For the RuBisCO genes, two phylotypes were found, both belonging to Gammaproteobacteria. One of them was closely related to the previously known mytilid symbiont, the other, to a pogonophore symbiont, presumably a methanotrophic bacterium. One phylotype of particulate methane oxygenase genes was also revealed; this finding indicated the presence of a methanotrophic symbiont. Phylogenetic analysis of the pmoA placed this endosymbiont within the Gammaproteobacteria, in a cluster including the methanotrophic bacterial genus Methylobacter and other methanotrophic Bathymodiolus gill symbionts. These results provide evidence for the existence of two types of endosymbionts (thioautotrophic and methanotrophic) in the gills of B. azoricus and demonstrate that, apart from the phylogenetic analysis of 16S rRNA genes, parallel analysis of functional genes is essential.     

Accelerated Sulfur Cycle in Coastal Marine Sediment beneath Areas of Intensive Shellfish Aquaculture

Prokaryotes in marine sediments taken from two neighboring semienclosed bays (the Yamada and Kamaishi bays) at the Sanriku coast in Japan were investigated by the culture-independent molecular phylogenetic approach coupled with chemical and activity analyses. These two bays were chosen in terms of their similar hydrogeological and chemical characteristics but different usage modes; the Yamada bay has been used for intensive shellfish aquaculture, while the Kamaishi bay has a commercial port and is not used for aquaculture. Substantial differences were found in the phylogenetic composition of 16S rRNA gene clone libraries constructed for the Yamada and Kamaishi sediments. In the Yamada library, phylotypes affiliated with δ-Proteobacteria were the most abundant, and those affiliated with γ-Proteobacteria were the second-most abundant. In contrast, the Kamaishi library was occupied by phylotypes affiliated with Planctomycetes, γ-Proteobacteria, δ-Proteobacteria, and Crenarchaeota. In the γ-Proteobacteria, many Yamada phylotypes were related to free-living and symbiotic sulfur oxidizers, whereas the Kamaishi phylotype was related to the genus Pseudomonas. These results allowed us to hypothesize that sulfate-reducing and sulfur-oxidizing bacteria have become abundant in the Yamada sediment. This hypothesis was supported by quantitative competitive PCR (qcPCR) with group-specific primers. The qcPCR also suggested that organisms closely related to Desulfotalea in the Desulfobulbaceae were the major sulfate-reducing bacteria in these sediments. In addition, potential sulfate reduction and sulfur oxidation rates in the sediment samples were determined, indicating that the sulfur cycle has become active in the Yamada sediment beneath the areas of intensive shellfish aquaculture.     

Evolutionary Dynamics of Ralstonia solanacearum

We investigated the genetic diversity, extent of recombination, natural selection, and population divergence of Ralstonia solanacearum samples obtained from sources worldwide. This plant pathogen causes bacterial wilt in many crops and constitutes a serious threat to agricultural production due to its very wide host range and aggressiveness. Five housekeeping genes, dispersed around the chromosome, and three virulence-related genes, located on the megaplasmid, were sequenced from 58 strains belonging to the four major phylogenetic clusters (phylotypes). Whereas genetic variation is high and consistent for all housekeeping loci studied, virulence-related gene sequences are more diverse. Phylogenetic and statistical analyses suggest that this organism is a highly diverse bacterial species containing four major, deeply separated evolutionary lineages (phylotypes I to IV) and a weaker subdivision of phylotype II into two subgroups. Analysis of molecular variations showed that the geographic isolation and spatial distance have been the significant determinants of genetic variation between phylotypes. R. solanacearum displays high clonality for housekeeping genes in all phylotypes (except phylotype III) and significant levels of recombination for the virulence-related egl and hrpB genes, which are limited mainly to phylotype strains III and IV. Finally, genes essential for species survival are under purifying selection, and those directly involved in pathogenesis might be under diversifying selection.     

Changes in Bacterioplankton Composition under Different Phytoplankton Regimens

The results of empirical studies have revealed links between phytoplankton and bacterioplankton, such as the frequent correlation between chlorophyll a and bulk bacterial abundance and production. Nevertheless, little is known about possible links at the level of specific taxonomic groups. To investigate this issue, seawater microcosm experiments were performed in the northwestern Mediterranean Sea. Turbulence was used as a noninvasive means to induce phytoplankton blooms dominated by different algae. Microcosms exposed to turbulence became dominated by diatoms, while small phytoflagellates gained importance under still conditions. Denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments showed that changes in phytoplankton community composition were followed by shifts in bacterioplankton community composition, both as changes in the presence or absence of distinct bacterial phylotypes and as differences in the relative abundance of ubiquitous phylotypes. Sequencing of DGGE bands showed that four Roseobacter phylotypes were present in all microcosms. The microcosms with a higher proportion of phytoflagellates were characterized by four phylotypes of the Bacteroidetes phylum: two affiliated with the family Cryomorphaceae and two with the family Flavobacteriaceae. Two other Flavobacteriaceae phylotypes were characteristic of the diatom-dominated microcosms, together with one Alphaproteobacteria phylotype (Roseobacter) and one Gammaproteobacteria phylotype (Methylophaga). Phylogenetic analyses of published Bacteroidetes 16S rRNA gene sequences confirmed that members of the Flavobacteriaceae are remarkably responsive to phytoplankton blooms, indicating these bacteria could be particularly important in the processing of organic matter during such events. Our data suggest that quantitative and qualitative differences in phytoplankton species composition may lead to pronounced differences in bacterioplankton species composition.     

Cyanobacterial Diversity in Natural and Artificial Microbial Mats of Lake Fryxell (McMurdo Dry Valleys, Antarctica): a Morphological and Molecular Approach

Currently, there is no consensus concerning the geographic distribution and extent of endemism in Antarctic cyanobacteria. In this paper we describe the phenotypic and genotypic diversity of cyanobacteria in a field microbial mat sample from Lake Fryxell and in an artificial cold-adapted sample cultured in a benthic gradient chamber (BGC) by using an inoculum from the same mat. Light microscopy and molecular tools, including 16S rRNA gene clone libraries, denaturing gradient gel electrophoresis, and sequencing, were used. For the first time in the study of cyanobacterial diversity of environmental samples, internal transcribed spacer (ITS) sequences were retrieved and analyzed to complement the information obtained from the 16S rRNA gene. Microscopy allowed eight morphotypes to be identified, only one of which is likely to be an Antarctic endemic morphotype. Molecular analysis, however, revealed an entirely different pattern. A much higher number of phylotypes (15 phylotypes) was found, but no sequences from Nodularia and Hydrocoryne, as observed by microscopy, were retrieved. The 16S rRNA gene sequences determined in this study were distributed in 11 phylogenetic lineages, 3 of which were exclusively Antarctic and 2 of which were novel. Collectively, these Antarctic sequences together with all the other polar sequences were distributed in 22 lineages, 9 of which were exclusively Antarctic, including the 2 novel lineages observed in this study. The cultured BGC mat had lower diversity than the field mat. However, the two samples shared three morphotypes and three phylotypes. Moreover, the BGC mat allowed enrichment of one additional phylotype. ITS sequence analysis revealed a complex signal that was difficult to interpret. Finally, this study provided evidence of molecular diversity of cyanobacteria in Antarctica that is much greater than the diversity currently known based on traditional microscopic analysis. Furthermore, Antarctic endemic species were more abundant than was estimated on the basis of morphological features. Decisive arguments concerning the global geographic distribution of cyanobacteria should therefore incorporate data obtained with the molecular tools described here.     

Novel Clade of Rickettsia spp. from Leeches

Intracellular rickettsia-like structures were found in the tissues of a glossiphoniid leech, Torix tagoi, by transmission electron microscopy. Diagnostic PCR analysis using specific primers suggested that of the nine glossiphoniid species examined, two species, T. tagoi and Hemicrepsis marginata, harbored bacteria of the genus Rickettsia. A 1.5-kb eubacterial 16S rRNA gene segment obtained from each of these species was amplified by PCR, cloned, and sequenced. Phylogenetic analysis of the 16S rRNA gene demonstrated that the Rickettsia species found in the leeches constituted a novel clade that is distinct from the clade of arthropod-associated Rickettsia species. In natural populations, 97.7% (43 of 44) of T. tagoi leeches and 100% (9 of 9) of H. marginata leeches carried Rickettsia, suggesting that infection with Rickettsia is prevalent in these leeches. This is the first report of Rickettsia found in annelids.

     

Species diversity of magnetotactic bacteria from the Ol’khovka River, Russia

A fraction of magnetotactic bacteria was isolated by magnetic separation from the water and silt samples collected from the Ol’khovka River (Kislovodsk, Russia). A 16S rRNA clone library was obtained from the total DNA of the fraction by PCR amplification and molecular cloning. Phylogenetic analysis of 67 16S rRNA gene sequences of randomly selected clones demonstrated that two phylotypes of magnetotactic bacteria were present in the library: the first phylotype consisted of 42 sequences and the second one included only one sequence. The remaining 24 sequences belonged to non-magnetotactic bacteria. According to the results of phylogenetic analysis, both phylotypes were magnetotactic cocci; the predominant sequences were almost identical to the 16S rRNA sequence of the freshwater coccus TB24 (X81185.1) identified earlier among the magnetotactic bacteria isolated from Lake Chiemsee (Bavaria). The phylotype represented by a single sequence formed a separate branch in the dendrogram, with 97% similarity between its sequence and that of TB24. The discovered phylotypes formed with the sequences of uncultured freshwater magnetotactic cocci a separate branch within the class Alphaproteobacteria and presumably belonged to a separate family within the recently described order Magnetococcales. Despite the fact that phylogenetic analysis of the 16S rRNA clone library did not reveal any phylotypes of magnetotactic spirilla, after the secondary enrichment of the fraction of magnetotactic bacteria using the “race track” technique, a new strain of magnetotactic spirilla, Magnetospirillum SO-1, was isolated. The closest relative of strain SO-1 was the previously described magnetotactic spirillum Magnetospirillum magneticum AMB-1.