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1.
Environ Microbiol ; 19(6): 2495-2506, 2017 06.
Artículo en Inglés | MEDLINE | ID: mdl-28464419

RESUMEN

Chemolithoautotrophic sulfur-oxidizing and denitrifying Gamma- (particularly the SUP05 cluster) and Epsilonproteobacteria (predominantly Sulfurimonas subgroup GD17) are assumed to compete for substrates (electron donors and acceptors) in marine pelagic redox gradients. To elucidate their ecological niche separation we performed 34 S0 , 15 NO3- and H13 CO3- stable-isotope incubations with water samples from Baltic Sea suboxic, chemocline and sulfidic zones followed by combined phylogenetic staining and high-resolution secondary ion mass spectrometry of single cells. SUP05 cells were small-sized (0.06-0.09 µm3 ) and most abundant in low-sulfidic to suboxic zones, whereas Sulfurimonas GD17 cells were significantly larger (0.26-0.61 µm3 ) and most abundant at the chemocline and below. Together, SUP05 and GD17 cells accumulated up to 48% of the labelled substrates but calculation of cell volume-specific rates revealed that GD17 cells incorporated labelled substrates significantly faster throughout the redox zone, thereby potentially outcompeting SUP05 especially at high substrate concentrations. Thus, in synopsis with earlier described features of SUP05/GD17 we conclude that their spatially overlapping association in stratified sulfidic zones is facilitated by their different lifestyles: whereas SUP05 cells are streamlined, non-motile K-strategists adapted to low substrate concentrations, GD17 cells are motile r-strategists well adapted to fluctuating substrate and redox conditions.


Asunto(s)
Crecimiento Quimioautotrófico/fisiología , Epsilonproteobacteria/crecimiento & desarrollo , Azufre/metabolismo , Desnitrificación , Epsilonproteobacteria/clasificación , Marcaje Isotópico , Océanos y Mares , Oxidación-Reducción , Oxígeno , Filogenia , Agua de Mar/microbiología
2.
Mar Genomics ; 24 Pt 3: 293-5, 2015 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-26476690

RESUMEN

A Vibrio sp. lytic phage VH7D was isolated from seawater of an abalone farm in Xiamen, China. The phage was capable of lysing Vibrio rotiferianus DSM 17186(T) and Vibrio harveyi DSM 19623(T). The complete genome of this phage consists of 246,964 nucleotides with a GC content of 41.31%, which characterized it as a giant vibriophage. Here we report the complete genome sequence and major findings from the genomic annotation.


Asunto(s)
Bacteriófagos/genética , Genoma Viral , ARN Viral/genética , Vibrio/virología , Bacteriófagos/clasificación , Regulación Viral de la Expresión Génica , Datos de Secuencia Molecular , Especificidad de la Especie , Proteínas Virales/genética , Proteínas Virales/metabolismo
3.
FEMS Microbiol Ecol ; 87(3): 770-9, 2014 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-24279499

RESUMEN

Pelagic redoxclines of the central Baltic Sea are dominated by the epsilonproteobacterial group Sulfurimonas GD17, considered to be the major driver of chemolithoautotrophic denitrification in this habitat. Autecological investigations of a recently isolated representative of this environmental group, Sulfurimonas gotlandica str. GD1(T), demonstrated that the bacterium grows best under sulfur-oxidizing, denitrifying conditions. However, in the presence of bicarbonate, this strain is able to use pyruvate as both an additional carbon source and an alternative electron donor. These observations suggested that the environmental group GD17 actively metabolizes organic substrates in situ. To examine this possibility, we used RNA-based stable isotope probing (RNA-SIP) on a natural redoxcline community provided with ¹³C-labeled pyruvate. While in this experiment, we were able to identify putative heterotrophic microorganisms, the uptake of ¹³C-pyruvate in GD17 nucleic acids could not be established. To resolve these contradictory findings, combined incorporation experiments with ¹4C- and ¹³C-labeled pyruvate were carried out in cells of strain GD1(T) cultivated under chemolithoautotrophic conditions, which favor pyruvate uptake rather than oxidation. An analysis of the labeled biomolecules revealed that pyruvate was mostly incorporated in cellular components such as amino acids, whose synthesis requires only minimal transformation. Carbon transfer into nucleic acids was not observed, explaining the inability of RNA-SIP to detect pyruvate incorporation by strain GD1(T) and the environmental group GD17. Together, these findings suggest that by integrating organic compounds such as pyruvate into cellular components S. gotlandica GD1(T) is able to replenish chemolithoautotrophic growth and thus ensure its survival in nutrient-limited habitats such as marine pelagic redoxclines.


Asunto(s)
Crecimiento Quimioautotrófico , Epsilonproteobacteria/metabolismo , Ácido Pirúvico/metabolismo , Agua de Mar/microbiología , Aminoácidos/química , Isótopos de Carbono/análisis , Desnitrificación , Ecosistema , Epsilonproteobacteria/crecimiento & desarrollo , Epsilonproteobacteria/aislamiento & purificación , Ácidos Grasos/química , Ácidos Nucleicos/química , Oxidación-Reducción
4.
Microbiologyopen ; 3(1): 80-8, 2014 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-24376054

RESUMEN

Epsilonproteobacteria have been found globally distributed in marine anoxic/sulfidic areas mediating relevant transformations within the sulfur and nitrogen cycles. In the Baltic Sea redox zones, chemoautotrophic epsilonproteobacteria mainly belong to the Sulfurimonas gotlandica GD17 cluster for which recently a representative strain, S. gotlandica GD1(T), could be established as a model organism. In this study, the potential effects of changes in dissolved inorganic carbon (DIC) and pH on S. gotlandica GD1(T) were examined. Bacterial cell abundance within a broad range of DIC concentrations and pH values were monitored and substrate utilization was determined. The results showed that the DIC saturation concentration for achieving maximal cell numbers was already reached at 800 µmol L(-1), which is well below in situ DIC levels. The pH optimum was between 6.6 and 8.0. Within a pH range of 6.6-7.1 there was no significant difference in substrate utilization; however, at lower pH values maximum cell abundance decreased sharply and cell-specific substrate consumption increased.


Asunto(s)
Carbono/farmacología , Crecimiento Quimioautotrófico/efectos de los fármacos , Epsilonproteobacteria/efectos de los fármacos , Concentración de Iones de Hidrógeno , Carga Bacteriana , Técnicas Bacteriológicas , Técnicas de Cultivo Celular por Lotes , Epsilonproteobacteria/crecimiento & desarrollo , Epsilonproteobacteria/metabolismo
5.
FEMS Microbiol Lett ; 348(2): 112-9, 2013 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-24102707

RESUMEN

Vibrio owensii is a potential bacterial pathogen in marine aquaculture system. In this study, five lytic phages specific against Vibrio strain B8D, closely related to V. owensii, were isolated from seawater of an abalone farm. The phages were characterized with respect to morphology, genome size, growth phenotype, as well as thermal, and pH stability. All phages were found to belong to the family Siphoviridae with long noncontractile tails and terminal fibers. Restriction analysis indicated that the five phages were dsDNA viruses with molecular weights ranging from c. 30 to 48 kb. One-step growth experiments revealed that the phages were heterogeneous in latent periods (10-70 min), rise periods (40-70 min), and burst sizes [23-331 plaque-forming units (PFU) per infected cell] at the same host strain. All phages were thermal stable and were tolerant to a wide range of pH. The results indicated that these phages could be potential candidates of a phage cocktail for biological control of V. owensii in aquaculture systems.


Asunto(s)
Bacteriófagos/aislamiento & purificación , Bacteriófagos/patogenicidad , Siphoviridae/aislamiento & purificación , Siphoviridae/patogenicidad , Vibrio/virología , Bacteriófagos/química , Bacteriófagos/genética , Genoma Viral , Calor , Concentración de Iones de Hidrógeno , Siphoviridae/química , Siphoviridae/genética
6.
Int J Syst Evol Microbiol ; 63(Pt 11): 4141-4148, 2013 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-23749282

RESUMEN

A psychro- and aerotolerant bacterium was isolated from the sulfidic water of a pelagic redox zone of the central Baltic Sea. The slightly curved rod- or spiral-shaped cells were motile by one polar flagellum or two bipolar flagella. Growth was chemolithoautotrophic, with nitrate or nitrite as electron acceptor and either a variety of sulfur species of different oxidation states or hydrogen as electron donor. Although the bacterium was able to utilize organic substances such as acetate, pyruvate, peptone and yeast extract for growth, these compounds yielded considerably lower cell numbers than obtained with reduced sulfur or hydrogen; in addition, bicarbonate supplementation was necessary. The cells also had an absolute requirement for NaCl. Optimal growth occurred at 15 °C and at pH 6.6-8.0. The predominant fatty acid of this organism was 16 : 1ω7c, with 3-OH 14 : 0, 16 : 0, 16 : 1ω5c+t and 18 : 1ω7c present in smaller amounts. The DNA G+C content was 33.6 mol%. As determined in 16S rRNA gene sequence phylogeny analysis, the isolate belongs to the genus Sulfurimonas, within the class Epsilonproteobacteria, with 93.7 to 94.2 % similarity to the other species of the genus Sulfurimonas, Sulfurimonas autotrophica, Sulfurimonas paralvinellae and Sulfurimonas denitrificans. However, the distinct physiological and genotypic differences from these previously described taxa support the description of a novel species, Sulfurimonas gotlandica sp. nov. The type strain is GD1(T) ( = DSM 19862(T) = JCM 16533(T)). Our results also justify an emended description of the genus Sulfurimonas.


Asunto(s)
Crecimiento Quimioautotrófico , Epsilonproteobacteria/clasificación , Filogenia , Agua de Mar/microbiología , Composición de Base , ADN Bacteriano/genética , Epsilonproteobacteria/genética , Epsilonproteobacteria/aislamiento & purificación , Ácidos Grasos/química , Hidrógeno/metabolismo , Datos de Secuencia Molecular , Nitratos/metabolismo , Nitritos/metabolismo , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Azufre/metabolismo , Microbiología del Agua
7.
ISME J ; 7(7): 1413-23, 2013 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-23446833

RESUMEN

The Baltic Sea receives large nitrogen inputs by diazotrophic (N2-fixing) heterocystous cyanobacteria but the significance of heterotrophic N2 fixation has not been studied. Here, the diversity, abundance and transcription of the nifH fragment of the nitrogenase enzyme in two basins of the Baltic Sea proper was examined. N2 fixation was measured at the surface (5 m) and in anoxic water (200 m). Vertical sampling profiles of >10 and <10 µm size fractions were collected in 2007, 2008 and 2011 at the Gotland Deep and in 2011 in the Bornholm Basin. Both of these stations are characterized by permanently anoxic bottom water. The 454-pyrosequencing nifH analysis revealed a diverse assemblage of nifH genes related to alpha-, beta- and gammaproteobacteria (nifH cluster I) and anaerobic bacteria (nifH cluster III) at and below the chemocline. Abundances of genes and transcripts of seven diazotrophic phylotypes were investigated using quantitative polymerase chain reaction revealing abundances of heterotrophic nifH phylotypes of up to 2.1 × 10(7) nifH copies l(-1). Abundant nifH transcripts (up to 3.2 × 10(4) transcripts l(-1)) within nifH cluster III and co-occurring N2 fixation (0.44±0.26 nmol l(-1) day(-1)) in deep water suggests that heterotrophic diazotrophs are fixing N2 in anoxic ammonium-rich waters. Our results reveal that N2 fixation in the Baltic Sea is not limited to illuminated N-deplete surface waters and suggest that N2 fixation could also be of importance in other suboxic regions of the world's oceans.


Asunto(s)
Bacterias/enzimología , Bacterias/genética , Biodiversidad , Oxidorreductasas/genética , Agua de Mar/microbiología , Bacterias/clasificación , Bacterias/metabolismo , Cianobacterias/enzimología , Cianobacterias/genética , Cianobacterias/metabolismo , Fijación del Nitrógeno/genética , Océanos y Mares , Filogenia , Agua de Mar/química
8.
FEMS Microbiol Ecol ; 85(2): 251-61, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-23521397

RESUMEN

Pelagic redoxclines represent chemical gradients of elevated microbial activities. While chemolithoautotrophic microorganisms in these systems are well known as catalysts of major biogeochemical cycles, comparable knowledge on heterotrophic organisms is scarce. Thus, in this study, identity and biogeochemical involvement of active heterotrophs were investigated in stimulation experiments and activity measurements based on samples collected from pelagic redoxclines of the central Baltic Sea in 2005 and 2009. In the 2009 samples, (13)C-acetate 16S rRNA stable isotope probing (16S rRNA-SIP) identified gammaproteobacteria affiliated with Colwellia sp. and Neptunomonas sp. in addition to epsilonproteobacteria related to Arcobacter spp. as active heterotrophs at the oxic-anoxic interface layer. Incubations from sulfidic waters were dominated by two phylogenetic subgroups of Arcobacter. In the 2005 samples, organics, manganese(IV), and iron(III) were added to the sulfidic waters, followed by the determination of metal reduction and identification of the stimulated organisms. Here, the same Arcobacter and Colwellia subgroups were stimulated as in 2009, with Arcobacter predominating in samples, in which manganese(IV) reduction was highest. Our results offer new insights into the heterotrophic bacterial assemblage of Baltic Sea pelagic redoxclines and suggest Arcobacter spp. as a heterotroph with presumed relevance also for manganese cycling.


Asunto(s)
Ácido Acético/metabolismo , Epsilonproteobacteria/metabolismo , Gammaproteobacteria/metabolismo , Procesos Heterotróficos , Agua de Mar/microbiología , Arcobacter/clasificación , Arcobacter/aislamiento & purificación , Arcobacter/metabolismo , Bacterias/clasificación , Bacterias/aislamiento & purificación , Bacterias/metabolismo , Secuencia de Bases , Epsilonproteobacteria/clasificación , Epsilonproteobacteria/aislamiento & purificación , Compuestos Férricos/metabolismo , Gammaproteobacteria/clasificación , Gammaproteobacteria/aislamiento & purificación , Manganeso/metabolismo , Datos de Secuencia Molecular , Océanos y Mares , Filogenia , ARN Ribosómico 16S/genética , Agua de Mar/química
9.
Environ Microbiol ; 15(5): 1505-13, 2013 May.
Artículo en Inglés | MEDLINE | ID: mdl-23013279

RESUMEN

Pelagic marine oxygen-depleted zones often exhibit a redox gradient, caused by oxygen depletion due to biological demand exceeding ventilation, and the accumulation of reduced chemical species, such as hydrogen sulfide. These redox gradients harbour a distinct assemblage of epsilonproteobacteria capable of fixing carbon dioxide autotrophically in the dark and potentially of utilizing hydrogen sulfide chemolithotrophically by oxidation with nitrate. Together, these two processes are referred to as chemolithoautotrophic denitrification. The focus of this study was the recently isolated and cultivated representative strain of pelagic epsilonproteobacteria, 'Sulfurimonas gotlandica' strain GD1, specifically dark carbon dioxide fixation and its substrate turnovers during chemolithotrophic denitrification. By connecting these processes stoichiometrically and comparing the results with those obtained for dark carbon dioxide fixation and nutrient concentrations measured in pelagic redox gradients of the Baltic Sea, we were able to estimate the role of chemolithoautotrophic denitrification in the environment. Evidence is provided for a defined zone where chemolithoautotrophic denitrification of these epsilonproteobacteria allows the complete removal of nitrate and hydrogen sulfide from the water column. This water layer is roughly equivalent in thickness to the average overlapping region of the two substrates, but slightly larger. Such a difference may be explained by a variety of reasons, including, e.g. utilization of substrates present at concentrations below the detection limit, alternative usage of other substrates as thiosulfate or nitrous oxide, or comparable activities of other microbes. However, the combined results of in vitro and in situ studies strongly suggest that epsilonproteobacteria are primarily responsible for hydrogen sulfide and nitrate removal from pelagic Baltic Sea redox gradients.


Asunto(s)
Crecimiento Quimioautotrófico/fisiología , Desnitrificación , Epsilonproteobacteria/metabolismo , Agua de Mar/microbiología , Dióxido de Carbono/metabolismo , Epsilonproteobacteria/aislamiento & purificación , Sulfuro de Hidrógeno/metabolismo , Nitratos/metabolismo , Océanos y Mares , Oxidación-Reducción , Oxígeno/metabolismo
10.
ISME J ; 6(2): 461-70, 2012 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-21776032

RESUMEN

An analysis of the microbial metabolism is fundamental to understanding globally important element transformations. One culture-independent approach to deduce those prokaryotic metabolic functions is to analyze metatranscriptomes. Unfortunately, since mRNA is extremely labile, it is unclear whether the abundance patterns detected in nature are vulnerable to considerable modification in situ simply due to sampling procedures. Exemplified on comparisons of metatranscriptomes retrieved from pelagic suboxic zones of the central Baltic Sea (70-120 m depth), earlier identified as areas of high aerobic ammonium oxidation activity, and quantification of specific transcripts in them, we show that different sampling techniques significantly influence the relative abundance of transcripts presumably diagnostic of the habitat. In situ fixation using our newly developed automatic flow injection sampler resulted in an abundance of thaumarchaeal ammonia monooxygenase transcripts that was up to 30-fold higher than that detected in samples obtained using standard oceanographic sampling systems. By contrast, the abundance of transcripts indicative of cellular stress was significantly greater in non-fixed samples. Thus, the importance of in situ fixation in the reliable evaluation of distinct microbial activities in the ecosystem based on metatranscriptomics is obvious. In consequence, our data indicate that the significance of thaumarchaeota to aerobic ammonium oxidation could yet have been considerably underestimated. Taken these results, this could in general also be the case in attempts aimed at an unbiased gene expression analysis of areas below the epipelagic zone, which cover 90% of the world's oceans.


Asunto(s)
Perfilación de la Expresión Génica/métodos , Agua de Mar/microbiología , Archaea/enzimología , Archaea/genética , Archaea/metabolismo , Países Bálticos , Ecosistema , Océanos y Mares , Oxidorreductasas/genética , Compuestos de Amonio Cuaternario/metabolismo , Transcriptoma
11.
Proc Natl Acad Sci U S A ; 109(2): 506-10, 2012 Jan 10.
Artículo en Inglés | MEDLINE | ID: mdl-22203982

RESUMEN

Eutrophication and global climate change lead to expansion of hypoxia in the ocean, often accompanied by the production of hydrogen sulfide, which is toxic to higher organisms. Chemoautotrophic bacteria are thought to buffer against increased sulfide concentrations by oxidizing hydrogen sulfide before its diffusion to oxygenated surface waters. Model organisms from such environments have not been readily available, which has contributed to a poor understanding of these microbes. We present here a detailed study of "Sulfurimonas gotlandica" str. GD1, an Epsilonproteobacterium isolated from the Baltic Sea oxic-anoxic interface, where it plays a key role in nitrogen and sulfur cycling. Whole-genome analysis and laboratory experiments revealed a high metabolic flexibility, suggesting a considerable capacity for adaptation to variable redox conditions. S. gotlandica str. GD1 was shown to grow chemolithoautotrophically by coupling denitrification with oxidation of reduced sulfur compounds and dark CO(2) fixation. Metabolic versatility was further suggested by the use of a range of different electron donors and acceptors and organic carbon sources. The number of genes involved in signal transduction and metabolic pathways exceeds those of other Epsilonproteobacteria. Oxygen tolerance and environmental-sensing systems combined with chemotactic responses enable this organism to thrive successfully in marine oxygen-depletion zones. We propose that S. gotlandica str. GD1 will serve as a model organism in investigations that will lead to a better understanding how members of the Epsilonproteobacteria are able to cope with water column anoxia and the role these microorganisms play in the detoxification of sulfidic waters.


Asunto(s)
Adaptación Fisiológica/fisiología , Epsilonproteobacteria/crecimiento & desarrollo , Epsilonproteobacteria/genética , Genoma Bacteriano/genética , Sulfuro de Hidrógeno/metabolismo , Anaerobiosis , Secuencia de Bases , Dióxido de Carbono/metabolismo , Citometría de Flujo , Genómica/métodos , Alemania , Redes y Vías Metabólicas/genética , Modelos Teóricos , Anotación de Secuencia Molecular , Datos de Secuencia Molecular , Océanos y Mares , Oxidación-Reducción , Análisis de Secuencia de ADN , Transducción de Señal/genética , Especificidad de la Especie
12.
FEMS Microbiol Ecol ; 74(1): 32-41, 2010 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-20649907

RESUMEN

Marine pelagic redoxclines are characterized by pronounced activities of chemolithoautotrophic microorganisms. As evidenced by the high dark CO(2) fixation rates measured around the oxic-anoxic interface but also in the upper sulfidic zone, the accordant organisms participate in important biogeochemical transformations. Although Epsilonproteobacteria have been identified as an important chemoautotrophic group in these environments, detailed species-level information on the identity of actively involved prokaryotes is lacking. In the present study, active chemolithoautotrophic prokaryotic assemblages were identified in the sulfidic zone of a pelagic Black Sea redoxcline by applying rRNA-based stable isotope probing in combination with 16S rRNA gene single-strand conformation polymorphism analysis and 16S rRNA gene cloning. The results showed that a single epsilonproteobacterium, affiliated with the genus Sulfurimonas, and two different members of the gammaproteobacterial sulfur oxidizer (GSO) cluster were responsible for dark CO(2) fixation activities in the upper sulfidic layer of the Black Sea redoxcline. Phylogenetically, these organisms were closely related to microorganisms, distributed worldwide, that are thought to be key players in denitrification and sulfide oxidation. Together, these findings emphasize the importance of chemolithoautotrophic members of the Sulfurimonas and GSO groups in the carbon, nitrogen, and sulfur cycles of oxic-anoxic pelagic transition zones.


Asunto(s)
Biodiversidad , Crecimiento Quimioautotrófico , Epsilonproteobacteria/clasificación , Gammaproteobacteria/clasificación , Agua de Mar/microbiología , Mar Negro , Dióxido de Carbono/metabolismo , Dermatoglifia del ADN , ADN Bacteriano/genética , Epsilonproteobacteria/genética , Gammaproteobacteria/genética , Datos de Secuencia Molecular , Filogenia , Polimorfismo Conformacional Retorcido-Simple , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN
13.
FEMS Microbiol Ecol ; 71(2): 226-36, 2010 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-19925634

RESUMEN

Oxic-anoxic interfaces harbor significant numbers and activity of chemolithoautotrophic microorganisms, known to oxidize reduced sulfur or nitrogen species. However, measurements of in situ distribution of bulk carbon dioxide (CO(2)) assimilation rates and active autotrophic microorganisms have challenged the common concept that aerobic and denitrifying sulfur oxidizers are the predominant autotrophs in pelagic oxic-anoxic interfaces. Here, we provide a comparative investigation of nutrient, sulfur, and manganese chemistry, microbial biomass distribution, as well as CO(2) fixation at the pelagic redoxcline of the eastern Gotland Basin, Baltic Sea. Opposing gradients of oxygen, nitrate, and sulfide approached the detection limits at the chemocline at 204 m water depth. No overlap of oxygen or nitrate with sulfide was observed, whereas particulate manganese was detected down to 220 m. More than 70% of the bulk dark CO(2) assimilation, totaling 9.3 mmol C m(-2) day(-1), was found in the absence of oxygen, nitrite, and nitrate and could not be stimulated by their addition. Maximum fixation rates of up to 1.1 mumol C L(-1) day(-1) were surprisingly susceptible to altered redox potential or sulfide concentration. These results suggest that novel redox-sensitive pathways of microbial sulfide oxidation could account for a significant fraction of chemolithoautotrophic growth beneath pelagic chemoclines. A mechanism of coupled activity of sulfur-oxidizing and sulfur-reducing microorganisms is proposed.


Asunto(s)
Dióxido de Carbono/metabolismo , Epsilonproteobacteria/metabolismo , Gammaproteobacteria/metabolismo , Azufre/metabolismo , Microbiología del Agua , Anaerobiosis , Biomasa , Nitratos/análisis , Nitritos/análisis , Oxidación-Reducción , Oxígeno/análisis , Agua de Mar/química , Agua de Mar/microbiología
14.
Environ Microbiol ; 11(2): 326-37, 2009 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-18793316

RESUMEN

Marine pelagic redoxclines are zones of high dark CO(2) fixation rates, which can correspond up to 30% of the surface primary production. However, despite this significant contribution to the pelagic carbon cycle, the identity of most chemolithoautotrophic organisms is still unknown. Therefore, the aim of this study was to directly link the dark CO(2) fixation capacity of a pelagic redoxcline in the central Baltic Sea (Landsort Deep) with the identity of the main chemolithoautotrophs involved. Our approach was based on the analysis of natural carbon isotope signatures in fatty acid methyl esters (FAMEs) and on measurements of CO(2) incorporation in (13)C-bicarbonate pulse experiments. The incorporation of (13)C into chemolithoautotrophic cells was investigated by rRNA-based stable isotope probing (RNA-SIP) and FAME analysis after incubation for 24 and 72 h under in situ conditions. Our results demonstrated that fatty acids indicative of Proteobacteria were significantly enriched in (13)C slightly below the chemocline. RNA-SIP analyses revealed that two different Gammaproteobacteria and three closely related Epsilonproteobacteria of the Sulfurimonas cluster were active dark CO(2)-fixing microorganisms, with a time-dependent community shift between these groups. Labelling of Archaea was not detectable, but after 72 h of incubation the (13)C-label had been transferred to a potentially bacterivorous ciliate related to Euplotes sp. Thus, RNA-SIP provided direct evidence for the contribution of chemolithoautotrophic production to the microbial food web in this marine pelagic redoxcline, emphasizing the importance of dark CO(2)-fixing Proteobacteria within this habitat.


Asunto(s)
Isótopos de Carbono/metabolismo , Epsilonproteobacteria/clasificación , Epsilonproteobacteria/metabolismo , Cadena Alimentaria , Gammaproteobacteria/clasificación , Gammaproteobacteria/metabolismo , Agua de Mar/microbiología , Animales , Archaea/clasificación , Archaea/genética , Archaea/metabolismo , Biodiversidad , Dióxido de Carbono/metabolismo , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Epsilonproteobacteria/genética , Euplotes/clasificación , Euplotes/genética , Euplotes/metabolismo , Ácidos Grasos/química , Gammaproteobacteria/genética , Datos de Secuencia Molecular , ARN/química , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Coloración y Etiquetado
15.
Appl Environ Microbiol ; 74(24): 7546-51, 2008 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-18952879

RESUMEN

Recent studies have indicated that chemoautotrophic Epsilonproteobacteria might play an important role, especially as anaerobic or microaerophilic dark CO(2)-fixing organisms, in marine pelagic redoxclines. However, knowledge of their distribution and abundance as actively CO(2)-fixing microorganisms in pelagic redoxclines is still deficient. We determined the contribution of Epsilonproteobacteria to dark CO(2) fixation in the sulfidic areas of central Baltic Sea and Black Sea redoxclines by combining catalyzed reporter deposition-fluorescence in situ hybridization with microautoradiography using [(14)C]bicarbonate and compared it to the total prokaryotic chemoautotrophic activity. In absolute numbers, up to 3 x 10(5) (14)CO(2)-fixing prokaryotic cells ml(-1) were enumerated in the redoxcline of the central Baltic Sea and up to 9 x 10(4) (14)CO(2)-fixing cells ml(-1) were enumerated in the Black Sea redoxcline, corresponding to 29% and 12%, respectively, of total cell abundance. (14)CO(2)-incorporating cells belonged exclusively to the domain Bacteria. Among these, members of the Epsilonproteobacteria were approximately 70% of the cells in the central Baltic Sea and up to 100% in the Black Sea. For the Baltic Sea, the Sulfurimonas subgroup GD17, previously assumed to be involved in autotrophic denitrification, was the most dominant CO(2)-fixing group. In conclusion, Epsilonproteobacteria were found to be mainly responsible for chemoautotrophic activity in the dark CO(2) fixation maxima of the Black Sea and central Baltic Sea redoxclines. These Epsilonproteobacteria might be relevant in similar habitats of the world's oceans, where high dark CO(2) fixation rates have been measured.


Asunto(s)
Dióxido de Carbono/metabolismo , Epsilonproteobacteria/aislamiento & purificación , Epsilonproteobacteria/metabolismo , Agua de Mar/microbiología , Autorradiografía , Biodiversidad , Radioisótopos de Carbono/metabolismo , Recuento de Colonia Microbiana , Oscuridad , Hibridación Fluorescente in Situ/métodos
16.
Appl Environ Microbiol ; 74(14): 4398-404, 2008 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-18502937

RESUMEN

The abundance, vertical distribution, and diversity of aerobic anoxygenic phototrophic bacteria (AAP) were studied at four basins of the Baltic Sea. AAP were enumerated by infrared epifluorescence microscopy, and their diversity was analyzed by using pufM gene clone libraries. In addition, numbers of CFU containing the pufM gene were determined, and representative strains were isolated. Both approaches indicated that AAP reached maximal abundance in the euphotic zone. Maximal AAP abundance was 2.5 x 10(5) cells ml(-1) (11% of total prokaryotes) or 1.0 x 10(3) CFU ml(-1) (9 to 10% of total CFU). Environmental pufM clone sequences were grouped into 11 operational taxonomic units phylogenetically related to cultivated members of the Alpha-, Beta-, and Gammaproteobacteria. In spite of varying pufM compositions, five clones were present in all libraries. Of these, Jannaschia-related clones were always found in relative abundances representing 25 to 30% of the total AAP clones. The abundances of the other clones varied. Clones potentially affiliated with typical freshwater Betaproteobacteria sequences were present at three Baltic Sea stations, whereas clones grouping with Loktanella represented 40% of the total cell numbers in the Gotland Basin. For three alphaproteobacterial clones, probable pufM phylogenetic relationships were supported by 16S rRNA gene analyses of Baltic AAP isolates, which showed nearly identical pufM sequences. Our data indicate that the studied AAP assemblages represented a mixture of marine and freshwater taxa, thus characterizing the Baltic Sea as a "melting pot" of abundant, polyphyletic aerobic photoheterotrophic bacteria.


Asunto(s)
Bacterias Aerobias/clasificación , Bacterias Aerobias/aislamiento & purificación , Bacterioclorofila A/biosíntesis , Alphaproteobacteria/clasificación , Alphaproteobacteria/aislamiento & purificación , Proteínas Bacterianas/genética , Secuencia de Bases , Betaproteobacteria/clasificación , Betaproteobacteria/aislamiento & purificación , Recuento de Colonia Microbiana , Agua Dulce/microbiología , Gammaproteobacteria/clasificación , Gammaproteobacteria/aislamiento & purificación , Genes Bacterianos , Variación Genética , Datos de Secuencia Molecular , Océanos y Mares , Proteínas del Complejo del Centro de Reacción Fotosintética/genética , Fototropismo , Filogenia , Polimorfismo de Longitud del Fragmento de Restricción , ARN Bacteriano/aislamiento & purificación , ARN Ribosómico 16S/aislamiento & purificación , Agua de Mar/microbiología , Microbiología del Agua
17.
Appl Environ Microbiol ; 73(22): 7155-61, 2007 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-17921285

RESUMEN

Members of the class Epsilonproteobacteria are known to be of major importance in biogeochemical processes at oxic-anoxic interfaces. In pelagic redoxclines of the central Baltic Sea, an uncultured epsilonproteobacterium related to Sulfurimonas denitrificans was proposed to play a key role in chemolithotrophic denitrification (I. Brettar, M. Labrenz, S. Flavier, J. Bötel, H. Kuosa, R. Christen, and M. G. Höfle, Appl. Environ. Microbiol. 72:1364-1372, 2006). In order to determine the abundance, activity, and vertical distribution of this bacterium in high-resolution profiles, 16S rRNA cloning and catalyzed reporter deposition and fluorescence in situ hybridization (CARD-FISH) and quantitative PCR measurements were carried out. The results showed that 21% of the derived clone sequences, which in the present study were grouped together under the name GD17, had >99% similarity to the uncultured epsilonproteobacterium. A specific gene probe against GD17 (S-*-Sul-0090-a-A-18) was developed and used for enumeration by CARD-FISH. In different pelagic redoxclines sampled during August 2003, May 2005, and February 2006, GD17 cells were always detected from the lower oxic area to the sulfidic area. Maximal abundance was detected around the chemocline, where sulfide and nitrate concentrations were close to the detection limit. The highest GD17 numbers (2 x 10(5) cells ml(-1)), representing up to 15% of the total bacteria, were comparable to those reported for Epsilonproteobacteria in pelagic redoxclines of the Black Sea and the Cariaco Trench (X. Lin, S. G. Wakeham, I. F. Putnam, Y. M. Astor, M. I. Scranton, A. Y. Chistoserdov, and G. T. Taylor, Appl. Environ. Microbiol. 72:2679-2690, 2006). However, in the Baltic Sea redoxclines, Epsilonproteobacteria consisted nearly entirely of cells belonging to the distinct GD17 group. This suggested that GD17 was the best-adapted epsilonproteobacterium within this ecological niche.


Asunto(s)
Epsilonproteobacteria/genética , Agua de Mar/microbiología , Técnicas de Tipificación Bacteriana , Dióxido de Carbono/metabolismo , Epsilonproteobacteria/clasificación , Epsilonproteobacteria/crecimiento & desarrollo , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , ARN Ribosómico 16S/genética , Estaciones del Año , Análisis de Secuencia de ADN
18.
Appl Environ Microbiol ; 71(11): 6664-72, 2005 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-16269695

RESUMEN

Anaerobic or microaerophilic chemolithoautotrophic bacteria have been considered to be responsible for CO2 dark fixation in different pelagic redoxclines worldwide, but their involvement in redox processes is still not fully resolved. We investigated the impact of 17 different electron donor/acceptor combinations in water of pelagic redoxclines from the central Baltic Sea on the stimulation of bacterial CO2 dark fixation as well as on the development of chemolithoautotrophic populations. In situ, the highest CO2 dark fixation rates, ranging from 0.7 to 1.4 micromol liter(-1) day(-1), were measured directly below the redoxcline. In enrichment experiments, chemolithoautotrophic CO2 dark fixation was maximally stimulated by the addition of thiosulfate, reaching values of up to 9.7 micromol liter(-1) CO2 day(-1). Chemolithoautotrophic nitrate reduction proved to be an important process, with rates of up to 33.5 micromol liter(-1) NO3(-) day(-1). Reduction of Fe(III) or Mn(IV) was not detected; nevertheless, the presence of these potential electron acceptors influenced the development of stimulated microbial assemblages. Potential chemolithoautotrophic bacteria in the enrichment experiments were displayed on 16S ribosomal complementary DNA single-strand-conformation polymorphism fingerprints and identified by sequencing of excised bands. Sequences were closely related to chemolithoautotrophic Thiomicrospira psychrophila and Maorithyas hadalis gill symbiont (both Gammaproteobacteria) and to an uncultured nitrate-reducing Helicobacteraceae bacterium (Epsilonproteobacteria). Our data indicate that this Helicobacteraceae bacterium could be of general importance or even a key organism for autotrophic nitrate reduction in pelagic redoxclines.


Asunto(s)
Dióxido de Carbono/metabolismo , Oscuridad , Transporte de Electrón , Epsilonproteobacteria/crecimiento & desarrollo , Gammaproteobacteria/crecimiento & desarrollo , Agua de Mar/microbiología , ADN Bacteriano/análisis , ADN Ribosómico/análisis , Ecosistema , Epsilonproteobacteria/clasificación , Epsilonproteobacteria/genética , Epsilonproteobacteria/metabolismo , Gammaproteobacteria/clasificación , Gammaproteobacteria/genética , Gammaproteobacteria/metabolismo , Datos de Secuencia Molecular , Nitratos/metabolismo , Oxidación-Reducción , Reacción en Cadena de la Polimerasa , Polimorfismo Conformacional Retorcido-Simple , ARN Ribosómico 16S/genética , Tiosulfatos/metabolismo
19.
Appl Environ Microbiol ; 69(4): 2395-8, 2003 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-12676728

RESUMEN

An indigenous freshwater bacterium (Sphingomonas sp. strain B18) from Lake Plubetasee (Schleswig-Holstein, Germany) was used to isolate 44 phages from 13 very different freshwater and brackish habitats in distant geographic areas. This bacterial strain was very sensitive to a broad spectrum of phages from different aquatic environments. Phages isolated from geographically distant aquatic habitats, but also those from the same sample, were diverse with respect to morphology and restriction pattern. Some phages were widely distributed, while different types coexisted in the same sample. It was concluded that phages could be a major factor in shaping the structure of bacterial communities and maintaining a high bacterial diversity.


Asunto(s)
Bacteriófagos/clasificación , Bacteriófagos/aislamiento & purificación , Agua Dulce/virología , Agua de Mar/virología , Sphingomonas/virología , Bacteriófagos/ultraestructura , Agua Dulce/microbiología , Variación Genética , Geografía , Lisogenia , Datos de Secuencia Molecular , Myoviridae/clasificación , Myoviridae/aislamiento & purificación , Myoviridae/ultraestructura , Podoviridae/clasificación , Podoviridae/aislamiento & purificación , Podoviridae/ultraestructura , Siphoviridae/clasificación , Siphoviridae/aislamiento & purificación , Siphoviridae/ultraestructura , Sphingomonas/aislamiento & purificación , Ensayo de Placa Viral
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