TY - GEN A1 - Böckelmann, Uta A1 - Janke, Andrea A1 - Kuhn, Ramona A1 - Neu, Thomas R. A1 - Wecke, Jörg A1 - Lawrence, John R. A1 - Szewzyk, Ulrich T1 - Bacterial extracellular DNA forming a defined network-like structure Y1 - 2006 ER - TY - GEN A1 - Guiot, Serge R. A1 - Kuhn, Ramona A1 - Lévesque, M. J. A1 - Cimpoia, Ruxandra T1 - Ultrastructure of a bioelectrolytic methanogenic/methanotrophic granular biofilm for the complete degradation of tetrachloroethylene in contaminated groundwater Y1 - 2007 ER - TY - GEN A1 - Guiot, Serge R. A1 - Cimpoia, Ruxandra A1 - Kuhn, Ramona A1 - Alaplantive, Aude T1 - Electrolytic Methanogenic-methanotropic coupling for Tetrachloroethylene Bioremediation: A Proof of Concept T2 - Environmental science & technology Y1 - 2008 SN - 0013-936X VL - 42 IS - 8 SP - 3011 EP - 3017 ER - TY - GEN A1 - Kuhn, Ramona A1 - Tóth, Elisabeth A1 - Geppert, Helmut A1 - Fischer, Thomas A1 - Martienssen, Marion T1 - Identification of the Complete Photodegradation Pathway of Ethylenediaminetetra(methylenephosphonic acid) in Aqueous Solution T2 - Clean - soil, air, water : a journal of sustainability and environmental safety N2 - This study aims at investigating the abiotic degradation pathway of ethylenediaminetetra(methylenephosphonic acid) (EDTMP) simulated applying UV irradiation. The degradation of EDTMP and formation of degradation products was determined using LC-MS and ³¹P-NMR. In the laboratory scale experiments, EDTMP was degraded within 30 min and the degradation products, iminodi(methylenephosphonic acid) (IDMP), ethylaminobis(methylenephosphonic acid) (EABMP), and amino- (methylenephosphonic acid) (AMPA), were simultaneously released. IDMP was the main degradation product of EDTMP. Therefore, we conclude that the initial cleavage of EDTMP is a heterolytically driven process, which starts the degradation process at the intramolecular C-N bond. In contrast, the main product of a possible homolytic C-C cleavage of methylaminobis(methylenephosphonic acid) could not be confirmed with either LC-MS or ³¹P-NMR. Additionally, there was no evidence for a primary attack on the C-P bond. All identified degradation products of EDTMP have been mineralized to carbon dioxide (CO₂). Three additional degradation products (M1, M2, and M3) have been found using the ³¹P-NMR analysis but have not yet been quantified using LC-MS. We assume that the unidentified degradation product M1 is related to m/z 312, M2 to m/z 341, and M3 to m/z 409. Thus we concluded that EDTMP undergoes photochemical conversion to IDMP, the main degradation product. EABMP and AMPA also accumulate, but in smaller amounts. All intermediates are further mineralized to CO₂. KW - EDTMP KW - LC-MS KW - 31P-NMR KW - Phosphonates KW - Photodegradation Y1 - 2017 U6 - https://doi.org/10.1002/clen.201500774 SN - 1863-0669 VL - 45 IS - 5 SP - 1 EP - 8 ER - TY - GEN A1 - Kuhn, Ramona A1 - Böllmann, Jörg A1 - Krahl, Kathrin A1 - Bryant, Isaac Mbir A1 - Martienssen, Marion T1 - Comparison of ten different DNA extraction procedures with respect to their suitability for environmental samples T2 - Journal of Microbiological Methods N2 - DNA extraction for molecular biological applications usually requires target optimized extraction procedures depending on the origin of the samples. For environmental samples, a range of different procedures has been developed. We compared the applicability and efficiency of ten selected DNA extraction methods published in recent literature using four different environmental samples namely: activated sludge from a domestic wastewater treatment plant, river sediment, anaerobic digestion sludge and nitrifying enrichment culture. We assessed the suitability of the extraction procedures based on both DNA yield and quality. DNA quantification was performed by both ultra violet (UV) spectrophotometry and fluorescence spectrophotometry after staining with PicoGreen. In our study, DNA yields based on UV measurement were overestimated in most cases while DNA yields from fluorescence measurements correlated well with the sample load on agarose gels of crude DNA. The quality of the DNA extracts was determined by gel electrophoresis of crude DNA and PCR products from 16S rDNA with the universal primer set 27f/1525r. It was observed that gel electrophoresis of crude DNA was not always suitable to evaluate DNA integrity and purity since interfering background substances (e.g. humic substances) were not visible. Therefore, we strongly recommend examining the DNA quality of both crude DNA and 16S rDNA PCR products by gel electrophoresis when a new extraction method is established. Summarizing, we found four out of ten extraction procedures being applicable to all tested samples without noticeable restrictions. The procedure G (according to the standard method 432_10401 of the Lower Saxony State Office for Consumer Protection and Food Safety) had the broadest application range over procedure J (published by Wilson, 2001). These were followed by procedures F (Singka et al., 2012) and A (Bourrain et al., 1999). All four extraction procedures delivered reliable and reproducible crude DNA and PCR products. From an economical point of view, all procedures tested during this study were cheaper compared to commercial DNA extraction kits. KW - DNA extraction KW - DNA quantification KW - DNA yield KW - Environmental samples KW - Gel electrophoresis KW - PicoGreen Y1 - 2017 U6 - https://doi.org/10.1016/j.mimet.2017.10.007 SN - 0167-7012 VL - 143 SP - 78 EP - 86 ER - TY - GEN A1 - Kuhn, Ramona A1 - Böllmann, Jörg A1 - Krahl, Kathrin A1 - Bryant, Isaac Mbir A1 - Martienssen, Marion T1 - Data on DNA gel sample load, gel electrophoresis, PCR and cost analysis T2 - Data in Brief N2 - The data presented in this article provide supporting information to the related research article “Comparison of ten different DNA extraction procedures with respect to their suitability for environmental samples” (revised manuscript submitted to J. Microbiol. Methods). In that article, we compared the suitability of ten selected DNA extraction methods based on DNA quality, purity, quantity and applicability to universal PCR. Here we provide the data on the specific DNA gel sample load, all unreported gel images of crude DNA and PCR results, and the complete cost analysis for all tested extraction procedures and in addition two commercial DNA extraction kits for soil and water. KW - Cost analysis KW - DNA sample load KW - Gel electrophoresis Y1 - 2018 U6 - https://doi.org/10.1016/j.dib.2017.11.082 SN - 2352-3409 VL - Vol. 16 SP - 732 EP - 751 ER - TY - CHAP A1 - Kuhn, Ramona A1 - Pollice, Alfieri A1 - Laera, Guiseppe A1 - Lipollis, Rosa A1 - Papa, Sergio ED - Lesjean, Boris T1 - Standard assays and metaproteomes as new approaches for functional characterization of membrane bioreactor biomass T2 - Membrane Technologies for Wastewater Treatment and Reuse, Proceedings der 2nd IWA National Young Water Professionals Conference in Berlin, 4.-6. Juni 2007 KW - Membrane bioreactors KW - biomass characterisation KW - EPS/SMP KW - metaproteomes Y1 - 2007 SN - 987-3-9811684-0-2 SP - 57 EP - 64 PB - IWA CY - Berlin ER - TY - GEN A1 - Kuhn, Ramona A1 - Benndorf, Dirk A1 - Rapp, Erdmann A1 - Reichl, Udo A1 - Palese, Luigi Leonardo A1 - Pollice, Alfieri T1 - Metaproteome analysis of sewage sludge from membrane bioreactors T2 - Proteomics N2 - Microbial dynamics and enzymatic activities of activated sludge processes are not completely understood yet. A better understanding about the biology is indispensable for further process optimization. Since proteins play a key role as catalysts in sludge processes, a protocol for protein extraction and analysis by 2-D PAGE was established. It is based on phenol extraction of alkaline extracts and on a subsequent precipitation with ammonium sulphate. 2-D protein patterns obtained from different sludges collected from membrane bioreactors showed – besides common spots – significant differences. Selected proteins were identified with nano- HPLC-ESI-MS/MS. All membrane biological reactor (MBR) sludge samples investigated in this study contained elastase 3A, which implies that this human serine protease is a significant constituent of municipal wastewater. Although the identification of proteins from ammonia-oxidizing bacterium Nitrosomonas europaea was expected, the detection of a protein with homology to the marine bacterium Saprospira grandis in MBR1 was surprising. KW - De novo sequencing KW - Membrane bioreactor KW - metaproteomics KW - microbial communities KW - Microbiology KW - Protein extraction Y1 - 2011 U6 - https://doi.org/10.1002/pmic.201000590 SN - 1615-9861 VL - 11 IS - 13 SP - 2738 EP - 2744 ER - TY - GEN A1 - Domańska, Magdalena A1 - Kuhn, Ramona A1 - Łomotowski, Janusz A1 - Stańczyk, Ewa T1 - FISH method for identification of microbes in wastewater distribution systems T2 - Environmental Protection Engineering N2 - The FISH (fluorescence in situ hybridization) method is widely used to identify various types of cells. In comparison to cultivation-dependent methods, identification of microbes by FISH is easier and generally takes several hours. This paper presents a review of improvements to the FISH method, its advantages and disadvantages, as well as examples of applications. Particular consideration was given to the efficiency of microbes identification in samples taken from sewerage. The effectiveness of the method was confirmed by the results obtained in samples from the membrane bioreactor (MBR). Y1 - 2014 SN - 0324-8828 VL - 40 IS - 3 SP - 151 EP - 160 ER - TY - GEN A1 - Kuhn, Ramona A1 - Jensch, Robert A1 - Bryant, Isaac Mbir A1 - Fischer, Thomas A1 - Liebsch, Stephan A1 - Martienssen, Marion T1 - The influence of selected bivalent metal ions on the photolysis of diethylenetriamine penta(methylenephosphonic acid) T2 - Chemosphere N2 - DTPMP is predominantly utilized as scale inhibitor. We investigated the reaction rates and degradation mechanism of DTPMP with and without addition of Fe²⁺, Mg²⁺ and Ca²⁺ by performing LC/MS and ³¹P-NMR analyses. DTPMP undergoes conversion with and without addition of bivalent metal ions. The initial cleavage of DTPMP is initiated at the C-N bond leading to release of IDMP as its major breakdown product. The release of smaller quantities of EABMP and AMPA confirmed the nucleophilic attack on the DTPMP amines. Oxidation of Fe²⁺ to Fe³⁺ during the initial 30 min indicated an intramolecular electron transfer changing the electron density distribution at the nitrogen centre, which increased the radical attack during UV irradiation. Independent of the fact that Fe acted as catalyst and Mg²⁺ and Ca²⁺ acted as reactants, we found no significant differences in their degradation mechanisms. However, the reaction rates were strongly affected by the addition of the bivalent metal ions as Fe²⁺ accelerated most DTPMP degradation followed by Mg²⁺ and Ca²⁺. The UV treatment without metal ion addition was four times slower compared with Fe²⁺ addition. We conclude that in environments rich in ferrous iron and/or at reduced redox potential, photolysis of DTPMP will be catalysed by iron and will lead to accumulation of IDMP, EABMP and AMPA and several other none-quantifiable breakdown products. KW - DTPMP KW - Ferrous ion KW - LC/MS KW - Phosphonates KW - Photolysis Y1 - 2018 U6 - https://doi.org/10.1016/j.chemosphere.2018.07.033 SN - 0045-6535 IS - 210 SP - 726 EP - 733 ER -