TY - JOUR A1 - Hartman, Jan F. A1 - Gentz, Torben A1 - Schiller, Amanda A1 - Greule, Markus A1 - Grossart, Hans-Peter A1 - Ionescu, Danny A1 - Keppler, Frank A1 - Martinez-Cruz, Karla A1 - Sepulveda-Jauregui, Armando A1 - Isenbeck-Schroeter, Margot T1 - A f ast and sensitive method for the continuous in situ determination of dissolved methane and its delta C-13-isotope ratio in surface waters JF - Limnology and Oceanography-methods N2 - A fast and sensitive method for the continuous determination of methane (CH4) and its stable carbon isotopic values (delta C-13-CH4) in surface waters was developed by applying a vacuum to a gas/liquid exchange membrane and measuring the extracted gases by a portable cavity ring-down spectroscopy analyser (M-CRDS). The M-CRDS was calibrated and characterized for CH4 concentration and delta C-13-CH4 with synthetic water standards. The detection limit of the M-CRDS for the simultaneous determination of CH4 and delta C-13-CH4 is 3.6 nmol L-1 CH4. A measurement precision of CH4 concentrations and delta C-13-CH4 in the range of 1.1%, respectively, 1.7 parts per thousand (1 sigma) and accuracy (1.3%, respectively, 0.8 parts per thousand [1 sigma]) was achieved for single measurements and averaging times of 10 min. The response time tau of 57 +/- 5 s allow determination of delta C-13-CH4 values more than twice as fast than other methods. The demonstrated M-CRDS method was applied and tested for Lake Stechlin (Germany) and compared with the headspace-gas chromatography and fast membrane CH4 concentration methods. Maximum CH4 concentrations (577 nmol L-1) and lightest delta C-13-CH4 (-35.2 parts per thousand) were found around the thermocline in depth profile measurements. The M-CRDS-method was in good agreement with other methods. Temporal variations in CH4 concentration and delta C-13-CH4 obtained in 24 h measurements indicate either local methane production/oxidation or physical variations in the thermocline. Therefore, these results illustrate the need of fast and sensitive analyses to achieve a better understanding of different mechanisms and pathways of CH4 formation in aquatic environments. Y1 - 2018 U6 - https://doi.org/10.1002/lom3.10244 SN - 1541-5856 VL - 16 IS - 5 SP - 273 EP - 285 PB - Wiley CY - Hoboken ER - TY - JOUR A1 - Cuadrat, Rafael R. C. A1 - Ionescu, Danny A1 - Davila, Alberto M. R. A1 - Grossart, Hans-Peter T1 - Recovering genomics clusters of secondary metabolites from lakes using genome-resolved metagenomics JF - Frontiers in microbiology N2 - Metagenomic approaches became increasingly popular in the past decades due to decreasing costs of DNA sequencing and bioinformatics development. So far, however, the recovery of long genes coding for secondary metabolites still represents a big challenge. Often, the quality of metagenome assemblies is poor, especially in environments with a high microbial diversity where sequence coverage is low and complexity of natural communities high. Recently, new and improved algorithms for binning environmental reads and contigs have been developed to overcome such limitations. Some of these algorithms use a similarity detection approach to classify the obtained reads into taxonomical units and to assemble draft genomes. This approach, however, is quite limited since it can classify exclusively sequences similar to those available (and well classified) in the databases. In this work, we used draft genomes from Lake Stechlin, north-eastern Germany, recovered by MetaBat, an efficient binning tool that integrates empirical probabilistic distances of genome abundance, and tetranucleotide frequency for accurate metagenome binning. These genomes were screened for secondary metabolism genes, such as polyketide synthases (PKS) and non-ribosomal peptide synthases (NRPS), using the Anti-SMASH and NAPDOS workflows. With this approach we were able to identify 243 secondary metabolite clusters from 121 genomes recovered from our lake samples. A total of 18 NRPS, 19 PKS, and 3 hybrid PKS/NRPS clusters were found. In addition, it was possible to predict the partial structure of several secondary metabolite clusters allowing for taxonomical classifications and phylogenetic inferences. Our approach revealed a high potential to recover and study secondary metabolites genes from any aquatic ecosystem. KW - metagenomics 2.0 KW - PKS KW - NRPS KW - freshwater KW - environmental genomics Y1 - 2018 U6 - https://doi.org/10.3389/fmicb.2018.00251 SN - 1664-302X VL - 9 PB - Frontiers Research Foundation CY - Lausanne ER - TY - JOUR A1 - Bizic-Ionescu, Mina A1 - Ionescu, Danny A1 - Grossart, Hans-Peter T1 - Organic Particles: Heterogeneous Hubs for Microbial Interactions in Aquatic Ecosystems JF - Frontiers in microbiology N2 - The dynamics and activities of microbes colonizing organic particles (hereafter particles) greatly determine the efficiency of the aquatic carbon pump. Current understanding is that particle composition, structure and surface properties, determined mostly by the forming organisms and organic matter, dictate initial microbial colonization and the subsequent rapid succession events taking place as organic matter lability and nutrient content change with microbial degradation. We applied a transcriptomic approach to assess the role of stochastic events on initial microbial colonization of particles. Furthermore, we asked whether gene expression corroborates rapid changes in carbon-quality. Commonly used size fractionated filtration averages thousands of particles of different sizes, sources, and ages. To overcome this drawback, we used replicate samples consisting each of 3–4 particles of identical source and age and further evaluated the consequences of averaging 10–1000s of particles. Using flow-through rolling tanks we conducted long-term experiments at near in situ conditions minimizing the biasing effects of closed incubation approaches often referred to as “the bottle-effect.” In our open flow-through rolling tank system, however, active microbial communities were highly heterogeneous despite an identical particle source, suggesting random initial colonization. Contrasting previous reports using closed incubation systems, expression of carbon utilization genes didn’t change after 1 week of incubation. Consequently, we suggest that in nature, changes in particle-associated community related to carbon availability are much slower (days to weeks) due to constant supply of labile, easily degradable organic matter. Initial, random particle colonization seems to be subsequently altered by multiple organismic interactions shaping microbial community interactions and functional dynamics. Comparative analysis of thousands particles pooled togethers as well as pooled samples suggests that mechanistic studies of microbial dynamics should be done on single particles. The observed microbial heterogeneity and inter-organismic interactions may have important implications for evolution and biogeochemistry in aquatic systems. KW - particle-associated bacteria KW - microbial communities KW - inter- and intra-species interactions KW - antagonism KW - phage KW - transcriptome Y1 - 2018 U6 - https://doi.org/10.3389/fmicb.2018.02569 SN - 1664-302X VL - 9 PB - Frontiers Research Foundation CY - Lausanne ER -