RESUMO
OBJECTIVE: The objectivewas to study the effect of changing the anglconvergence of the abutment walls, depending on their height, as well as the number of supports, on the accuracy of fitting metal-ceramic prostheses to the abutments. MATERIAL AND METHODS: It was studied 72 constructions: prostheses on single abutments with abutment heights from 3 mm to 9 mm with an increase in height of 2 mm, an angle of convergence of the abutment walls from 0° to 10° with an increase in convergence of 2° in the amount of 24 prostheses; 24 framework designs supported by two abutments, with the same parameters (height and angle of inclination of the abutment walls) and prostheses supported by three abutments in the amount of 24 designs. The abutments were milled from a titanium alloy Ti-6Al-4V, and the prostheses were cast from a cobalt-chrome alloy. During fabrication of the prostheses, a compensation varnish was applied to the respective abutments to create a gap between the framework and the abutment for a 50 µm thick cement film. The crown prostheses were cemented onto the abutments with GC Fuji + glass-ionomer cement, followed by vertical sawing in the medio-distal direction. After sawing, the gaps between the inner edges of the prostheses walls and the outer edges of the abutment walls were measured using a «Levenhuk DTX 90¼ digital microscope (Russia) at x100 magnification. RESULTS: Abatement height 3 mm with any number of supports and 5 mm with up to 3 supports do not need taper for precise fitting. In 5 mm abatements and 3 and more supports minimal required taper is 2°. Seven mm singular abatement requires 4° taper, but the increased number of supports enhances it up to 6°. For 9 mm abatement optimal abatement taper for the best fitting is 8°. CONCLUSION: The study showed the relationship between the fitting accuracy and the change in the abutment taper, their height, and the number of supports in the structure.
Assuntos
Dente Suporte , Implantes Dentários , Coroas , Cimentos Dentários , Materiais Dentários , Prótese Dentária Fixada por Implante , Análise do Estresse Dentário , HumanosRESUMO
A novel quinazoline series of photoswitchable combretastatin A-4 (CA-4) analogues were synthesized and their photochemical properties and antiproliferative activity against A431 epidermoid carcinoma cells were studied. It was found that quinazoline analogues, in contrast to the majority of the known CA-4, exhibit high antiproliferative activity in the E-form as well. Photoswitching of the E-form to the Z-form resulted in a multiple (9-fold) increase in antiproliferative activity. 1H NMR monitoring showed that these compounds are very resistant to UV (λ = 365 nm) or sunlight irradiation and do not undergo photodegradation with a loss of antiproliferative activity that is inherent in heterocyclic analogues of CA-4. Similar photoswitching and an increase in antiproliferative activity are observed on exposure to sunlight. A selected compound (1a-Z51) in sub-micromolar concentrations induced apoptosis in A431 cells, while rad50/ATM/p53 were not involved in cell death. The growth of A431 cells was significantly inhibited after combination treatment with compound 1a-Z51 and chemotherapy drugs (cisplatin or 5-fluorouracil). In summary, the quinazoline analogues of CA-4 represent a promising strategy to achieve a photoswitchable potency for the treatment of cancers, including the development of combination therapies.
Assuntos
EstilbenosRESUMO
THE AIM OF THE STUDY: Was to evaluate the abrasive effect of toothpaste with detonation nanodiamonds synthesis of various concentrations on the enamel of human teeth. MATERIALS AND METHODS: The analysis of micrographs obtained by scanning electron microscopy showed a different abrasive and polishing effect of nanodiamonds in all the observed anatomical zones on the vestibular surface of the enamel of human teeth. RESULTS: No negative effect of the paste containing nanodiamonds on the enamel of human teeth was noted. At all concentrations of pastes containing nanodiamonds, on the vestibular surface of the enamel of the studied teeth, depot areas of densely attached particles of nanodiamonds were formed, a tendency was observed towards the formation of depots. The enamel of human teeth was mostly influenced by the concentration of nanodiamonds (2%, 2.5%). On the surface of the enamel were formed not only a depot of single nanodiamonds, but also accumulations were noted, merging into a layer in the form of a protective film. CONCLUSION: The study showed that toothpastes with nanoalases can reduce the enamel porosity, form a protective film on its surface, and thereby increase its microhardness and increase caries resistance.
Assuntos
Nanodiamantes , Dente , Esmalte Dentário , Humanos , Microscopia Eletrônica de Varredura , Cremes DentaisRESUMO
BACKGROUND: Due to the growth of oncological diseases of the maxillofacial region, there is an increasing need to improve methods of orthopedic treatment of post-resection defects of the upper jaw with modern designs of replacement prostheses. OBJECTIVE: Improving the quality of orthopedic treatment by creating an improved upper jaw resection prosthesis and justifying its use. MATERIAL AND METHODS: We studied the effectiveness of orthopedic treatment of post-resection defects of the upper jaw using the proposed replacement prosthesis with a pneumatic obturator. A special questionnaire was used to analyze the level of adaptation and maladaptation of patients. RESULTS: There was a significant decrease in complaints and adaptation time in the group of patients with a replacement prosthesis with a pneumatic obturator. CONCLUSION: The use of the proposed resection removable prosthesis helps to reduce the period of adaptation to the prosthesis, which indicates that it can be widely used in clinical practice.
Assuntos
Maxila , Obturadores Palatinos , Humanos , Maxila/cirurgiaRESUMO
Herein we present the synthesis of the two Lewis acids Al[N(C6F5)2]3 (ALTA) and Ga[N(C6F5)2]3 (GATA) via salt elimination reactions. The metal complexes were characterized by NMR-spectroscopic methods and X-ray diffraction analysis revealing the stabilization of the highly Lewis acidic metal centers by secondary metal-fluorine contacts. The Lewis acidic properties of Al[N(C6F5)2]3 and Ga[N(C6F5)2]3 are demonstrated by reactions with Lewis bases resulting in the formation of metallates accompanied by crucial structural changes. The two metallates [Cs(Tol)3]+[FAl(N(C6F5)2)3]- and [AsPh4]+[ClGa(N(C6F5)2)3]- contain interesting weakly coordinating anions. The reaction of Al[N(C6F5)2]3 with trityl fluoride yielded [CPh3]+[FAl(N(C6F5)2)3]- which could find application in the activation of metallocene polymerization catalysts. The qualitative Lewis acidity of Al[N(C6F5)2]3 and Ga[N(C6F5)2]3 was investigated by means of competition experiments for chloride ions in solution. DFT calculations yielded fluoride ion affinities in the gas phase (FIA) of 555 kJ mol-1 for Al[N(C6F5)2]3 and 472 kJ mol-1 for Ga[N(C6F5)2]3. Thus, Al[N(C6F5)2]3 can be considered a Lewis superacid with a fluoride affinity higher than SbF5 (493 kJ mol-1) whereas the FIA of the corresponding gallium complex is slightly below the threshold to Lewis superacidity.
RESUMO
Propionate can be directly oxidized anaerobically with sulfate as e-acceptor at haloalkaline conditions either incompletely to acetate (an example is Desulfobulbus alkaliphilus), or completely (for example by the members of genus Desulfonatronobacter). An enrichment with propionate at methanogenic conditions (without sulfate) inoculated with mixed sediments from hypersaline soda lakes in Kulunda Steppe (Altai, Russia) resulted in a domination of a new member of Syntrophobacteraceae (Deltaproteobacteria) in a consortium with the haloalkaliphilic lithotrophic methanogen Methanocalculus alkaliphilus. Transfer of this culture to a medium containing propionate as e-donor and sulfate as e-acceptor resulted in a disappearance of the methanogen and sulfide formation by the bacterial component, finally isolated into a pure culture at these conditions. Strain APr1 formed a distinct phylogenetic lineage within the family Syntrophobacteraceae, being equally distant from its members at the genus level. Phenotypically, strain APr1 resembled the species of the genus Syntrophobacter with substrate spectrum restricted to propionate and propanol utilized with sulfate, sulfite and thiosulfate as the e-acceptors. Propionate is oxidized incompletely to acetate. It is a moderately salt-tolerant (max. 1.2 M Na+) obligate alkaliphile (pH opt. 10). The isolate is proposed to be classified as a new candidate genus and species 'Candidatus Desulfonatronobulbus propionicus'.
Assuntos
Deltaproteobacteria/isolamento & purificação , Lagos/microbiologia , Tolerância ao Sal , Deltaproteobacteria/classificação , Deltaproteobacteria/genética , Deltaproteobacteria/metabolismo , Concentração de Íons de Hidrogênio , Filogenia , Propionatos/metabolismo , Sulfatos/metabolismoRESUMO
AIM: to compare the effectiveness of biomicroscopy (BMS) and optical coherence tomography (OCT) in recognizing prognostically unfavorable signs in peripheral vitreoretinal dystrophy (PVRD) patients. MATERIAL AND METHODS: A total of 131 cases of equatorial PVRD (91 eyes of 56 patients) were assessed. The mean patient's age was 24.7 years. The length of the anterior-posterior axis of the eyeball averaged 25.36±1.12 mm. Prevalence of particular warning signs in PVRD patients at BMS or OCT was comparatively analyzed. RESULTS: In 46 eyes with lattice dystrophy it was difficult to determine the presence of vitreoretinal traction at BMS; at OCT, areas of retinal adhesion to the posterior hyaloid membrane (PHM) along the edges of PVRD zones were revealed in all eyes. Of 31 eyes with «snail tracks¼ defects of the retina, 6 were diagnosed at BMS; in OCT scans of these patients, the PHM appeared firmly fixed along the edges of PVRD zones in all cases. As to horseshoe retinal tears (valve-like tears), BMS allowed to visualize vitreoretinal tractions in 7 of 12 eyes, while OCT revealed a tight contact between the PHM and the apex of the retinal flap in 11 of 12 eyes. In 7 eyes with retinoschisis we failed to detect any retinal traction at either BMS or OCT. In «non-differentiable¼ PVRD, BMS was also not able to reveal any vitreoretinal traction, while OCT was - in all 12 cases. CONCLUSION: OCT has proved much more effective than BMS in recognizing prognostically unfavorable signs in particular clinical forms of PVRD, such as vitreoretinal tractions, retinal defects, and intraretinal cavities.
Assuntos
Microscopia/métodos , Distrofias Retinianas , Tomografia de Coerência Óptica/métodos , Adolescente , Adulto , Pesquisa Comparativa da Efetividade , Feminino , Humanos , Masculino , Oftalmoscopia/métodos , Reprodutibilidade dos Testes , Distrofias Retinianas/classificação , Distrofias Retinianas/diagnóstico , Distrofias Retinianas/diagnóstico por imagem , Distrofias Retinianas/fisiopatologiaRESUMO
We studied the mechanisms of adaptation of human breast cancer cells MCF-7 to hypoxia and analyzed the role of AMPK/mTOR signaling pathway in the maintenance of cell proliferation under hypoxic conditions. It was found that long-term culturing (30 days or more) of MCF-7 cells under hypoxic conditions induced their partial adaptation to hypoxia. Cell adaptation to hypoxia was associated with attenuation of hypoxia-dependent AMPK induction with simultaneous constitutive activation of mTOR and Akt. These findings suggest that these proteins can be promising targets for targeted therapy of tumors developing under hypoxic conditions.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Adaptação Fisiológica/fisiologia , Neoplasias da Mama/metabolismo , Hipóxia Celular/fisiologia , Serina-Treonina Quinases TOR/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Ativação Enzimática , Feminino , Humanos , Células MCF-7RESUMO
Recent intensive microbiological investigation of sulfidogenesis in soda lakes did not result in isolation of any pure cultures of sulfate-reducing bacteria (SRB) able to directly oxidize acetate. The sulfate-dependent acetate oxidation at haloalkaline conditions has, so far, been only shown in two syntrophic associations of novel Syntrophobacteraceae members and haloalkaliphilic hydrogenotrophic SRB. In the course of investigation of one of them, obtained from a hypersaline soda lake in South-Western Siberia, a minor component was observed showing a close relation to Desulfonatronobacter acidivorans--a "complete oxidizing" SRB from soda lakes. This organism became dominant in a secondary enrichment with propionate as e-donor and sulfate as e-acceptor. A pure culture, strain APT3, was identified as a novel member of the family Desulfobacteraceae. It is an extremely salt-tolerant alkaliphile, growing with butyrate at salinity up to 4 M total Na(+) with a pH optimum at 9.5. It can grow with sulfate as e-acceptor with C3-C9 VFA and also with some alcohols. The most interesting property of strain APT3 is its ability to grow with acetate as e-donor, although not with sulfate, but with sulfite or thiosulfate as e-acceptors. The new isolate is proposed as a new species Desulfonatronobacter acetoxydans.
Assuntos
Deltaproteobacteria/isolamento & purificação , Lagos/microbiologia , Tolerância ao Sal , Sequência de Bases , Deltaproteobacteria/classificação , Deltaproteobacteria/genética , Deltaproteobacteria/metabolismo , Dados de Sequência Molecular , FilogeniaRESUMO
The composition and macroscopic structure of the floating oxygenic phototrophic communities from Kulunda steppe soda lakes (Petukhovskoe sodovoe, Tanatara VI, and Gorchiny 3) was described based on the data of the 2011 and 2012 expeditions (Winogradsky Institute of Microbiology). The algo-bacterial community with a green alga Ctenocladus circinnatus as an edificator was the typical one. Filamentous Geitlerinema sp. and Nodosilinea sp. were the dominant cyanobacteria. Apart from C. circinnatus, the algological component of the community contained unicellular green algae Dunaliella viridis and cf. Chlorella minutissima, as well as diatoms (Anomeoneis sphaerophora, Brchysira brebissonii, Brachysira zellensis, Mastogloia pusilla var. subcapitata, Nitzschia amphibia, Nitzschia communis, and Nitzschia sp.1). The latter have not been previously identified in the lakes under study. In all lakes, a considerable increase in salinity was found to result in changes in the composition and macroscopic structure of algo-bacterial communities.
Assuntos
Clorófitas/genética , Cianobactérias/genética , Diatomáceas/genética , Lagos/microbiologia , Filogenia , Microbiologia da Água , Clorófitas/classificação , Clorófitas/ultraestrutura , Cianobactérias/classificação , Cianobactérias/ultraestrutura , DNA de Algas/análise , DNA de Algas/genética , DNA Bacteriano/análise , DNA Bacteriano/genética , Diatomáceas/classificação , Diatomáceas/ultraestrutura , Pradaria , Lagos/química , Tipagem Molecular , RNA Ribossômico 16S/genética , RNA Ribossômico 18S/genética , Salinidade , SibériaRESUMO
The compensation of cell motion is an important step in single-particle tracking analysis of live cells. This step is required in most of the cases, since the movement of subcellular foci is superimposed by the movement and deformation of the cell, while only the local motion of the foci is important to be analysed. The cell motion and deformation compensation is usually performed by means of image registration. There are a number of approaches with different models and properties presented in the literature that perform cell image registration. However, the evaluation of the registration approach quality on real data is a tricky problem due to the fact that some stable features in the images with a priori no local motion are needed. In this paper we propose a methodology for creating live cell nuclei image sequences with stable features imposed. The features are defined using the regions of fluorescence bleaching invoked by the UV laser. Data with different deformations are acquired and can be used for evaluation of the cell image registration methods. Along with that, we describe an image analysis technique and a metric that can characterize the quality of the method quantitatively. The proposed methodology allows building a ground truth dataset for testing and thoroughly evaluating cell image registration methods.
Assuntos
Movimento Celular , Núcleo Celular/metabolismo , Algoritmos , Sobrevivência Celular , Bases de Dados como Assunto , Células HeLa , Humanos , Processamento de Imagem Assistida por ComputadorRESUMO
Protein arginine methyltransferases (PRMTs) are responsible for symmetric and asymmetric methylation of arginine residues of nuclear and cytoplasmic proteins. In the nucleus, PRMTs belong to important chromatin modifying enzymes of immense functional significance that affect gene expression, splicing and DNA repair. By time-lapse microscopy we have studied the sub-cellular localization and kinetics of PRMT1 after inhibition of PRMT1 and after irradiation. Both transiently expressed and endogenous PRMT1 accumulated in cytoplasmic bodies that were located in the proximity of the cell nucleus. The shape and number of these bodies were stable in untreated cells. However, when cell nuclei were microirradiated by UV-A, the mobility of PRMT1 cytoplasmic bodies increased, size was reduced, and disappeared within approximately 20 min. The same response occurred after γ-irradiation of the whole cell population, but with delayed kinetics. Treatment with PRMT1 inhibitors induced disintegration of these PRMT1 cytoplasmic bodies and prevented formation of 53BP1 nuclear bodies (NBs) that play a role during DNA damage repair. The formation of 53BP1 NBs was not influenced by PRMT1 overexpression. Taken together, we show that PRMT1 concentrates in cytoplasmic bodies, which respond to DNA injury in the cell nucleus, and to treatment with various PRMT1 inhibitors.
Assuntos
Citoplasma/enzimologia , Dano ao DNA , Raios gama , Proteína-Arginina N-Metiltransferases/metabolismo , Proteínas Repressoras/metabolismo , Raios Ultravioleta , Animais , Proteínas Cromossômicas não Histona/genética , Proteínas Cromossômicas não Histona/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Células HeLa , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Camundongos , Proteína-Arginina N-Metiltransferases/antagonistas & inibidores , Proteína-Arginina N-Metiltransferases/genética , Proteínas Repressoras/antagonistas & inibidores , Proteínas Repressoras/genética , Proteína 1 de Ligação à Proteína Supressora de Tumor p53RESUMO
Chitin is a dominant structural polymer produced in large amounts by brine shrimp Artemia in hypersaline lakes. Microbiological analysis of chitin utilization as a growth substrate in hypersaline chloride-sulfate lakes in the south Kulunda Steppe (Altai, Russia) revealed two groups of bacteria able to grow on chitin at moderate salinity. Under aerobic conditions, an enrichment culture was obtained at 2 M NaCl. Further purification resulted in the isolation of strains HCh1 and strain HCh2, identified as representatives of the genera Saccharospirillum and Arhodomonas (both in the Gammaproteobacteria). The chitin-utilizing potential has not been previously recognized in these genera. The Saccharospirillum sp. strain HCh1 grew on chitin within the salinity range from 0.5 to 3.25 M NaCl (optimum at 1 M), while Arhodomonas sp. strain HCh2 grew up to 2.5 M NaCl but had a higher salt optimum at 1.5 M. Anaerobic enrichments grew with chitin at 2 and 4 M NaCl, but growth in the latter was extremely slow and the culture eventually lost viability. The enrichment at 2 M NaCl resulted in the isolation of strain HCh-An1, identified as a distant new species of the genus Orenia in the clostridial order Halanaerobiales. It was able to grow on chitin within a salinity range from 1.0 to 2.5 M NaCl (optimum at 1.5 M). The strain is proposed as a new species of the genus Orenia-O. chitinitropha.
Assuntos
Quitina/metabolismo , Gammaproteobacteria/isolamento & purificação , Tolerância ao Sal , Sequência de Bases , Gammaproteobacteria/genética , Gammaproteobacteria/metabolismo , Lagos/química , Lagos/microbiologia , Dados de Sequência Molecular , RNA Ribossômico 16S/química , RNA Ribossômico 16S/genética , SalinidadeRESUMO
Various previously developed techniques for DNA extraction from the samples with complex physicochemical structure (soils, silts, and sediments) and modifications of these techniques developed in the present work were tested. Their usability for DNA extraction from the sediments of the Kulunda Steppe hypersaline soda lakes was assessed, and the most efficient procedure for indirect (two-stage) DNA extraction was proposed. Almost complete separation of the cell fraction was shown, as well as the inefficiency of nested PCR for analysis of the clone libraries obtained from washed sediments by amplification of the 16S rRNA gene fragments. Analysis of the clone library obtained from the cell fractions of stratified sediments (upper, medium, and lower layers) revealed that in the sediments of Lake Gorchina-3 most eubacterial phylotypes belonged to the class Clostridia, phylum Firmicutes. They were probably specific for this habitatand formed a new, presently unknown high-rank taxon. The data obtained revealed no pronounced stratification of the spe- cies diversity of the eubacterial component of the microbial community inhabiting the sediments (0-20 cm) in the inshore zone of Lake Gorchina-3.
Assuntos
Bactérias/genética , Sedimentos Geológicos/microbiologia , Filogenia , Bactérias/isolamento & purificação , Biodiversidade , DNA/isolamento & purificação , Biblioteca Gênica , Dados de Sequência Molecular , RNA Ribossômico 16S/genética , Sibéria , Microbiologia da ÁguaRESUMO
Phylogenetic analysis of the nifH genes, encoding the Fe protein of the nitrogenas enzymatic complex, was carried out for pure cultures of anoxygenic phototrophic bacteria of diverse origin, as well as for heterotrophic alkaliphilic sulfate reducers isolated from saline and soda lakes. Topology of the nitrogenase tree correlated with that of the 16S rRNAgene tree to a considerable degree; which niade it possible to use the nifH gene as a molecular marker for investigation of diazotrophic bacterialcommunities in silty sediments of saline and sodalakes. Although diazotrophs were revealed in all environmentalsamples, their phylogenetic diversity was relatively low. Sulfate-reducing deltaproteobacteria and photo- and chemotrophicgammaproteobacteria were predominant in samples integrated over sediment thickness. Analysis of samples fromthe upper sediment layers revealed predominance of phototrophic diazotrophs of various phyla, including purple sulfur and nonsulfur proteobacteria, green nonsulfur bacteria, heliobacteria; and cyanobacteria. Some phylotypes could not be identified, probably indicating the presence of bacterial groups which have not yet been studied by conventional microbiological techniques.
Assuntos
Bactérias/genética , Proteínas de Bactérias/genética , Oxirredutases/genética , Filogenia , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Microbiologia da Água , Bactérias/metabolismo , Proteínas de Bactérias/metabolismo , Marcadores Genéticos , Lagos/microbiologia , Oxirredutases/metabolismo , RNA Bacteriano/metabolismo , RNA Ribossômico 16S/metabolismoRESUMO
The complete genome sequence of a novel halophilic archaeon, Salinarchaeum sp. strain HArcht-Bsk1(T), was determined using next-generation sequencing. The genome comprises a 3,255,260-bp circular chromosome with a G+C content of 66.7%. Automatic annotation of the genome revealed a single rRNA operon, 45 tRNAs, and 3,013 protein-coding gene sequences.
RESUMO
Two obligately anaerobic sulfidogenic bacterial strains were isolated from the full-scale Thiopaq bioreactor in Lelystad (The Netherlands) removing H2S from biogas under oxygen-limiting and moderately haloalkaline conditions. Strain HSRB-L represents a dominant culturable sulfate-reducing bacterium in the reactor. It utilizes formate, H2 (with acetate as C-source) and lactate as e-donors, and sulfate, thiosulfate and sulfite as e-acceptors. It is haloalkalitolerant, with a pH range for lithotrophic growth from 7.5 to 9.7 (optimum at 8.5-9) and a salt range from 0.1 to 1.75 M total Na(+) (optimum at 0.6 M). The strain is a member of the genus Desulfonatronum and is proposed as a novel species D. alkalitolerans. The second strain, strain HTRB-L1, represents a dominant thiosulfate/sulfur reducer in the reactor. It is an obligate anaerobe utilizing formate and H2 (with acetate as C-source), lactate, pyruvate and fumarate as e-donors, and thiosulfate (incomplete reduction), sulfur, arsenate and fumarate as e-acceptors. With lactate as e-donor it also grows as an ammonifyer in the presence of nitrate and nitrite. HTRB-L1 is haloalkalitolerant, with a pH range for lithotrophic growth from 7.1 to 9.7 (optimum at 8.5) and a salt range from 0.6 to 1.5 M total Na(+) (optimum at 0.6 M). Phylogenetic analysis showed that strain HTRB-L1 is a novel species within the genus Sulfurospirillum (Epsilonproteobacteria) for which a name Sulfurospirillum alkalitolerans is proposed.
Assuntos
Reatores Biológicos/microbiologia , Deltaproteobacteria/isolamento & purificação , Epsilonproteobacteria/isolamento & purificação , Sequência de Bases , DNA Bacteriano/química , Deltaproteobacteria/genética , Deltaproteobacteria/metabolismo , Epsilonproteobacteria/genética , Epsilonproteobacteria/metabolismo , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA , Enxofre/metabolismoRESUMO
Chitin is produced in large amounts in hypersaline habitats with neutral pH due to the high biomass production of brine shrimp Artemia. Recently, a high abundance of Artemia was also noticed in hypersaline soda lakes in the Kulunda Steppe (Altai, Russia), which prompted us to survey the possibility of microbial chitin utilization at extremely haloalkaline conditions in soda brines. Most active chitin utilisation-supporting microbial growth was found at anaerobic conditions at pH 10 and up to 3.5 M total Na(+). At aerobic conditions, the degradation of chitin was slower, mostly incomplete and active at <2 M total Na(+), although very slow partial degradation was possible up to 4 M Na(+). Anaerobic enrichments at pH 10 yielded two different groups of obligately haloalkaliphilic fermentative anaerobes, exclusively specialized to utilise insoluble chitin as the only growth substrate. One group was represented by a single strain growing at moderate salinity, and another comprised multiple isolates growing up to 3.5 M Na(+). These groups represent two novel bacterial phyla not closely related to any other cultured bacteria. Aerobic enrichments from the lake sediments were dominated by several obligately haloalkaliphilic members of the genus Marinimicrobium in the Gammaproteobacteria. They were less specialised than the anaerobes and grew with chitin and its monomer and oligomers at a pH of 10 up to 2.5 M Na(+). Furthermore, several strains of haloalkaliphilic Gram-positive chitinolytics belonging to bacilli and actinobacteria were isolated from soda lake sediments and surrounding soda soils. In general, the results indicate the presence of an active and diverse haloalkaliphilic chitinolytic microbial community in hypersaline soda habitats.
Assuntos
Quitina/metabolismo , Gammaproteobacteria/isolamento & purificação , Gammaproteobacteria/metabolismo , Anaerobiose , Sequência de Bases , Ecossistema , Gammaproteobacteria/genética , Genes de RNAr , Sedimentos Geológicos/química , Sedimentos Geológicos/microbiologia , Concentração de Íons de Hidrogênio , Dados de Sequência Molecular , Filogenia , Tolerância ao Sal , Sibéria , SódioRESUMO
An anaerobic enrichment culture inoculated with a sample of sediments from soda lakes of the Kulunda Steppe with elemental sulfur as electron acceptor and formate as electron donor at pH 10 and moderate salinity inoculated with sediments from soda lakes in Kulunda Steppe (Altai, Russia) resulted in the domination of a Gram-positive, spore-forming bacterium strain AHT28. The isolate is an obligate anaerobe capable of respiratory growth using elemental sulfur, thiosulfate (incomplete reduction) and arsenate as electron acceptor with H2, formate, pyruvate and lactate as electron donor. Growth was possible within a pH range from 9 to 10.5 (optimum at pH 10) and a salt concentration at pH 10 from 0.2 to 2 M total Na+ (optimum at 0.6 M). According to the phylogenetic analysis, strain AHT28 represents a deep independent lineage within the order Bacillales with a maximum of 90 % 16S rRNA gene similarity to its closest cultured representatives. On the basis of its distinct phenotype and phylogeny, the novel haloalkaliphilic anaerobe is suggested as a new genus and species, Desulfuribacillus alkaliarsenatis (type strain AHT28(T) = DSM24608(T) = UNIQEM U855(T)).
Assuntos
Bacillales , Lagos/microbiologia , Microbiologia da Água , Arseniatos/metabolismo , Bacillales/classificação , Bacillales/citologia , Bacillales/genética , Bacillales/isolamento & purificação , Bacillales/metabolismo , Sequência de Bases , Concentração de Íons de Hidrogênio , Dados de Sequência Molecular , Oxirredução , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Sibéria , Enxofre/metabolismoRESUMO
Four strains of lithotrophic sulfate-reducing bacteria (SRB) have been enriched and isolated from anoxic sediments of hypersaline chloride-sulfate lakes in the Kulunda Steppe (Altai, Russia) at 2 M NaCl and pH 7.5. According to the 16S rRNA gene sequence analysis, the isolates were closely related to each other and belonged to the genus Desulfonatronovibrio, which, so far, included only obligately alkaliphilic members found exclusively in soda lakes. The isolates utilized formate, H(2) and pyruvate as electron donors and sulfate, sulfite and thiosulfate as electron acceptors. In contrast to the described species of the genus Desulfonatronovibrio, the salt lake isolates could only tolerate high pH (up to pH 9.4), while they grow optimally at a neutral pH. They belonged to the moderate halophiles growing between 0.2 and 2 M NaCl with an optimum at 0.5 M. On the basis of their distinct phenotype and phylogeny, the described halophilic SRB are proposed to form a novel species within the genus Desulfonatronovibrio, D. halophilus (type strain HTR1(T) = DSM24312(T) = UNIQEM U802(T)).