Vibrio spp. are ubiquitous in aquatic environments. In temperate regions, including the Baltic Sea, these bacteria, which include facultative pathogenic species, proliferate during warm water periods and may thus pose a risk to human health. However, while present at relatively high abundances in the summer months, Vibrio spp. are rarely detected during cold water periods. Several potential winter reservoirs, such as fish and sediment, have been discussed, but the seeding sources that give rise to Vibrio during the summer months are unknown. In this study, we employed a combination of cultivation-independent genus-specific digital droplet PCR, Illumina 16S rRNA gene sequencing, culture-dependent determinations of colony-forming units (CFU) and genusspecific strain isolation to identify possible winter reservoirs of Vibrio spp. Using these techniques, we demonstrate that in the southern Baltic Sea especially fish can serve as a winter reservoir for distinct Vibrio assemblages, including facultative pathogenic V. vulnificus. Particularly the invasive species round goby (Neogobius melanostomus), with its potential to appear in high individual densities, has the capacity to carry a comparable Vibrio spp. cell number like water and sediment, while only accounting for 1/1000 of the spatial share in the overall coastal environment. Thus, by providing a winter reservoir for distinct Vibrio communities, coastal fish and, especially areas in which round goby densities are high, may be hot spots of pathogenic Vibrio species.

The bacterial genus Vibrio is widespread in aquatic systems and contains several pathogenic species. As pathogenic Vibrio infections in aquaculture systems have been documented, the release of untreated aquaculture wastewater could cause the release of high doses of the pathogens into the environment and thus to the replacement of naturally occurring Vibrio populations. Aquaculture is one of the main industries on the island of Hainan (southern China) and the risk of pathogenic Vibrio contamination of the environment and humans is accordingly high. In this study, we used Illumina 16S rRNA gene sequencing of water, sediment, seagrass and tissue samples obtained from aquaculture operations in this region and from adjoining coastal and riverine areas to examine the impact of aquaculture on Vibrio assemblages. The results indicated that the effects of aquaculture on Vibrio community structure and abundance depended on the particular system. Classical pond aquaculture decreased Vibrio diversity and significantly reduced the relative abundances of Vibrio 16S rRNA genes compared to coastal reference populations. Aquaculture systems continuously flushed with marine water had a Vibrio community structure and diversity similar to that of coastal populations but with elevated levels of Vibrio sequence reads. Despite the observed changes, there was no clear evidence that aquaculture facilities altered Vibrio composition or led to a measurable increase in the relative abundance of Vibrio genes in the environment.

SummaryChemolithoautotrophic 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 34S0, 15 and H13 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.

Für MR Angiografie werden räumlich hochaufgelöste Mehrphasen-3D-(MP3D) und zunehmend auch zeitlich hochaufgelöste 4D-Techniken eingesetzt. Bisher existieren kaum systematische Analysen zur Optimierung von Kontrastmittel-Applikationen (KM). Ziel dieser Studie war, quantitative und qualitative Daten zur Applikation von Gadobutrol und Gadoterate Meglumine bei 1,5 und 3T zu erheben.

Vibrio harveyi and related vibrios are potential pathogens causing luminous vibriosis in marine aquaculture systems. In this study, two lytic phages P4A and P4F isolated using Vibrio strains B4A and B4F as indicator bacteria, respectively, were isolated from seawater of an abalone farm. Vibrio strain B4F belongs to the Harveyi clade of the genus Vibrio and was found to cause mortality of abalones in laboratory microcosms. Both phages were able to lyse Vibrio strain B4F. Electron microscopy revealed that phage P4A had an icosahedral head while P4F possessed an elongated hexagonal head. Both phages belong to the family Siphoviridae with long non-contractile tails. Restriction endonuclease analysis indicated that both phages were double-stranded DNA viruses and the genome sizes of P4A and P4F were estimated to be about 49 and 44 kb respectively. One-step growth curves revealed that these two phages exhibited distinct latent periods, exponential periods and burst sizes by infecting the same Vibrio strain B4F. Both phages were able to significantly reduce Vibrio population density in biofilm formed by Vibrio strain B4F on the surface of polyethylene film. It is suggested that these two phages may be promising candidates as biocontrol agents of infections caused by Vibrio strains belonging to the Haveyi clade in marine aquaculture systems.

The energy resolving capabilities of Photon Counting Detectors (PCD) in Computed Tomography (CT) facilitate energy-sensitive measurements. The provided image-information can be processed with Dual Energy and Multi Energy algorithms. A research PCD-CT firstly allows acquiring images with a close to clinical configuration of both the X-ray tube and the CT-detector. In this study, two algorithms (Material Decomposition and Virtual Non-Contrast-imaging (VNC)) are applied on a data set acquired from an anesthetized rabbit scanned using the PCD-CT system. Two contrast agents (CA) are applied: A gadolinium (Gd) based CA used to enhance contrasts for vascular imaging, and xenon (Xe) and air as a CA used to evaluate local ventilation of the animal's lung. Four different images are generated: a) A VNC image, suppressing any traces of the injected Gd imitating a native scan, b) a VNC image with a Gd-image as an overlay, where contrast enhancements in the vascular system are highlighted using colored labels, c) another VNC image with a Xe-image as an overlay, and d) a 3D rendered image of the animal's lung, filled with Xe, indicating local ventilation characteristics. All images are generated from two images based on energy bin information. It is shown that a modified version of a commercially available dual energy software framework is capable of providing images with diagnostic value obtained from the research PCD-CT system.

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.

Für die kontrastverstärkte MRA (CE-MRA) existieren zahlreiche Techniken sowie zahlreiche Publikationen, welche die Vor- und Nachteile der verschiedenen Techniken beleuchten. In Bezug auf die KM-Applikation ist die Datenlage jedoch sehr inhomogen. Ziel dieser Studie war die systematische Evaluation verschiedener KM-Applikationsschemata in Bezug auf die Konzentration, das Signal, sowie die Form der Boluskurve.

The influence of hydrodynamic events on the distribution of methane and its microbial turnover was investigated during the period from August 2011 to August 2013 along a transect from the eastern (EGB) to the western Gotland Basin (WGB), central Baltic Sea. The water column was characterized by a pronounced methane concentration gradient between the methane-rich deep anoxic and the methane-poor upper oxic water layer. In both basins, enhanced vertical turbulent diffusivities in fall (November 2011) and winter (February 2012) lead to an enhanced flux of methane from the deep anoxic water towards the oxic–anoxic transition zone (redox zone). In both basins, the increased vertical transport of methane in fall/winter was mirrored by reduced methane turnover times measured within the redox zone. Moreover, specific biomarkers indicative for aerobic methanotrophic bacteria implied an increase in the microbial population size from August 2011 till February 2012, indicating a methanotrophic community adapting to the variable methane fluxes. The deep water methane inventory of the EGB showed a seasonal pattern, with concentrations increasing during spring (May) and summer (August) and decreasing during fall (November) and winter (February) as a direct result of the seasonality of the vertical turbulent diffusivity. In contrast, the WGB showed no clear correlation between the seasons and the observed deep water methane variability. Here, the impact of lateral weak intrusions penetrating the deep water layer was identified as the main factor controlling the variability of the deep water methane concentration.Moreover, methane concentration and carbon stable isotopic data (δ13C CH4) demonstrate that the previously reported production of methane in the oxic water column below the thermocline occurs in the entire central Baltic Sea from May through November, and despite the large methane pool in the underlying anoxic deep water, might govern the moderate methane flux to the atmosphere in this area in summer.

A total of 19 bacterial strains were isolated from aquaculture water of an abalone farm in Xiamen.All these strains were identified as Vibrio spp.based on 16S rDNA gene sequence analysis.Their 16S rDNA sequences showed high degrees of similarity (≥98.99%)with closest matched reference sequences of Vibrio type species. However,due to extreme high similarity of 16S rDNA sequences among Vibrio species,it was impossible to identify these Vibrio strains to species level only based on 16S rDNA sequence analysis.Phylogenteic analysis also showed that most 16S rDNA sequences of these isolates were not grouped with the reference sequences of Vibrio type spe-cies,suggesting that 16S rDNA gene is not adequate for resolution of Vibrio species.Multilocus sequence analysis of 4 housekeeping genes (rpoA,pyrH,gapA,and topA)was further performed to identify these Vibrio isolates.The results showed that 19 isolates belong to two Vibrio species,Vibrio alginalyticus and Vibrio diabolicus,indicating that these two Vibrio species are dominant in aquaculture water of the abalone farm.Polymorphism analysis demon-strated that all of 4 housekeeping genes showed higher polymorphic sites than the 16S rDNA gene.The concatena-ted sequences of 4 housekeeping genes exhibited 41.1%of polymorphic sites,much higher than that of 16S rDNA (13.4%).These results suggested that the housekeeping genes showed remarkable higher solution than 16S rDNA gene for species identification of marine Vibrios.

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 μ mol 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.

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.

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.

Pelagic methane oxidation was investigated in dependence on differing hydrographic conditions within the redox zone of the Gotland Deep (GD) and Landsort Deep (LD), central Baltic Sea. The redox zone of both deeps, which indicates the transition between oxic and anoxic conditions, was characterized by a pronounced methane concentration gradient between the deep water (GD: 1233 nM, 223 m; LD: 2935 nM, 422 m) and the surface water (GD and LD < 10 nM). This gradient together with a 13C CH4 enrichment (δ13C CH4 deep water: GD −84‰, LD −71‰; redox zone: GD −60‰, LD −20‰; surface water: GD −47‰, LD −50‰; δ13C CH4 vs. Vienna Pee Dee Belemnite standard), clearly indicating microbial methane consumption within the redox zone. Expression analysis of the methane monooxygenase identified one active type I methanotrophic bacterium in both redox zones. In contrast, the turnover of methane within the redox zones showed strong differences between the two basins (GD: max. 0.12 nM d−1, LD: max. 0.61 nM d−1), with a nearly four-times-lower turnover time of methane in the LD (GD: 455 d, LD: 127 d). Vertical mixing rates for both deeps were calculated on the base of the methane concentration profile and the consumption of methane in the redox zone (GD: 2.5 × 10–6 m2 s−1, LD: 1.6 × 10–5 m2 s−1). Our study identified vertical transport of methane from the deep-water body towards the redox zone as well as differing hydrographic conditions (lateral intrusions and vertical mixing) within the redox zone of these deeps as major factors that determine the pelagic methane oxidation.

1. We describe the abundance of an autochthonous alphaproteobacteria Sphingomonas sp. and its lytic phages during a period of about 3 months in Lake Plussee, Northern Germany. This is the first report of a specific autochthonous phage host system of a heterotrophic bacterial strain in a natural freshwater environment. 2. The concentration of bacterial hosts (Sphingomonas sp. strain B18) fluctuated between 20 and 1,150 (median 168) colony-forming units (CFU) mL-1 and the concentration of the specific lytic phages ranged from 20 to 680 (median 110) plaque-forming units (PFU) mL-1. Different Sphingomonas sp. strains were isolated, showing different sensitivity against two phage isolates. The specific virus to bacteria ratio (VBR) varied by a factor of about 280 and ranged from 0.03 to about 8.5 (median 0.6). 3. Encounter rates of 0.0007 to 0.198 d-1 (median 0.03 d-1) were calculated from the measured concentrations of Sphingomonas sp. and its lytic phages. From changes of PFU between two consecutive samplings net phage decay rates in a range between 0.003 and 1.3 d-1 (median 0.26 d-1) were calculated. 4. Concentration factors were calculated which enable sufficient encounter rates to explain the concentrations of bacteria and their phages. 5. The results of Kokjohn et al. (1991) support the suggestion that the explanation of the coexistence of lytic phages and their host bacteria by the used encounter rate model needs to be revised.

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 GD1T ( = DSM 19862T = JCM 16533T). Our results also justify an emended description of the genus Sulfurimonas .

The Baltic Sea receives large nitrogen inputs by diazotrophic (N 2 -fixing) heterocystous cyanobacteria but the significance of heterotrophic N 2 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. N 2 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 N 2 fixation (0.44±0.26 nmol l −1 day −1 ) in deep water suggests that heterotrophic diazotrophs are fixing N 2 in anoxic ammonium-rich waters. Our results reveal that N 2 fixation in the Baltic Sea is not limited to illuminated N-deplete surface waters and suggest that N 2 fixation could also be of importance in other suboxic regions of the world’s oceans.

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.