TY - CONF A1 - Martin-Sanchez, Pedro Maria T1 - Fast and reliable qPCR-based detection of microbial contamination in fuel systems T2 - 5th International Symposium on Applied Microbiology and Molecular Biology in Oil Systems (ISMOSS) CY - Stavanger, Norway DA - 2015-06-02 PY - 2015 AN - OPUS4-33589 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Martin-Sanchez, Pedro Maria T1 - A novel qPCR protocol to the specific detection and quantification of Hormoconis resinae in fuel systems N2 - A novel qPCR protocol to the specific detection and quantification of Hormoconis resinae in fuel systems P.M. Martin-Sanchez1 , J. Toepel1, H.J. Kunte1, A.A. Gorbushina1, 2 1BAM Federal Institute for Materials Research and Testing, Department 4 Materials & Environment, Berlin, Germany 2 Free University of Berlin, Department of Earth Sciences & Biology, Chemistry & Pharmacy, Berlin, Germany Introduction. Microbial contamination of fuels is a phenomenon widely reported in the literature causing dramatic problems in fuel systems such as blockage of pipelines and filters, and corrosion of their metal components. A variety of microorganisms, fungi, yeasts and bacteria, can contaminate the distillate fuels. Between them, the filamentous fungus Hormoconis resinae, previously described as Cladosporium resinae and also known as "kerosene fungus" or "diesel bug", has traditionally been considered the main responsible of such troubles due to its great capacity to produce biomass and biofilms. Aim. The overarching goal of this study was to develop a novel real-time quantitative PCR (qPCR) method to detect and quantify the fungus H. resinae in samples from fuel systems. Materials and methods. Four specific primer sets targeting on two molecular markers, RNA polymerase II second largest subunit (RPB2) and internal transcribed spacers of rDNA (ITS), were designed using Primer Blast. The specificity of primer sets was evaluated by both conventional PCR and qPCR, analyzing the DNA extracts from H. resinae strains and other fungi and yeasts previously described in fuels as well as four taxonomically closest related species. The efficiency and sensitivity of these protocols were assessed by constructing the corresponding standard curves for each primer sets. Results. Two primer sets, Hr556F/Hr814R (RPB2) and Hr101F/Hr408R (ITS), were selected according to their remarkable specificity. Their standard curves showed a good correlation coefficient and efficiency. Additionally, the selected primer pairs showed a high sensitivity detecting a tiny amount of H. resinae DNA, 10 pg for RPB2 primers and 0.1 pg for ITS primers. Conclusions. The established qPCR protocol is a reliable and useful tool to the specific detection and quantification of H. resinae. Its subsequent implementation to analyze samples of fuels or biofilms covering fuel systems will allow the early detection of fungal outbreaks in order to apply the appropriated control procedures. Moreover, these analyses will determine the current incidence of this species in the cases of biological fuel contamination. Keywords. Hormoconis resinae, real-time qPCR, diesel bug, kerosene fungus T2 - XVI International Biodeterioration and Biodegradation Symposium; International Biodeterioration & Biodegradation Society CY - Lodz, Poland DA - 2014-09-03 PY - 2014 AN - OPUS4-32165 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Martin-Sanchez, Pedro Maria A1 - Gebhardt, Christopher A1 - Toepel, Jörg A1 - Barry, J. A1 - Munzke, N. A1 - Günster, Jens A1 - Gorbushina, Anna T1 - Monitoring microbial soiling in photovoltaic systems: A qPCR-based approach N2 - Soiling of photovoltaic (PV) systems compromises their performance causing a significant power loss and demanding periodical cleaning actions. This phenomenon raises great concerns in the solar energy field, thus leading to notable research efforts over the last decades. Soiling is caused by a dual action of dust deposition and biofouling. However, surprisingly, the microbiological contribution to PV soiling is often overlooked or underestimated. In this study, a variety of qPCR-based methods have been developed to quantify the microbial load of fungi, bacteria and phototrophs on PV panels. These protocols were evaluated by comparison with culturedependent methods, and were implemented with real solar plants for two years. The results show that the developed molecular methods are highly sensitive and reliable to monitor the microbial component of the soiling. Fungal biomass was clearly dominant in all analysed PV modules, while bacteria and phototrophs showed much lower abundance. Light microscopy and qPCR results revealed that melanised microcolonial fungi and phototrophs are the main biofilm-forming microorganisms on the studied solar panels. In particular, the fungal qPCR protocol is proposed as a useful tool for monitoring of PV soiling, and investigating the microbial contribution to specific soiling cases. KW - Solar panels KW - PV modules KW - Real-time qPCR KW - Bacteria KW - Fungi KW - Phototrophs PY - 2018 DO - https://doi.org/10.1016/j.ibiod.2017.12.008 SN - 0964-8305 VL - 129 SP - 13 EP - 22 PB - Elsevier Science AN - OPUS4-43892 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Martin-Sanchez, Pedro Maria A1 - Gorbushina, Anna A1 - Kunte, Hans-Jörg A1 - Toepel, Jörg T1 - A novel qPCR protocol for the specific detection and quantification of the fuel-deteriorating fungus Hormoconis resinae N2 - A wide variety of fungi and bacteria are known to contaminate fuels and fuel systems. These microbial contaminants have been linked to fuel system fouling and corrosion. The fungus Hormoconis resinae, a common jet fuel contaminant, is used in this study as a model for developing innovative risk assessment methods. A novel qPCR protocol to detect and quantify H. resinae in, and together with, total fungal contamination of fuel systems is reported. Two primer sets, targeting the markers RPB2 and ITS, were selected for their remarkable specificity and sensitivity. These primers were successfully applied on fungal cultures and diesel samples demonstrating the validity and reliability of the established qPCR protocol. This novel tool allows clarification of the current role of H. resinae in fuel contamination cases, as well as providing a technique to detect fungal outbreaks in fuel systems. This tool can be expanded to other well-known fuel-deteriorating microorganisms. KW - Microbial contamination KW - Real-time quantitative PCR KW - Microbiologically influenced corrosion; KW - Diesel biodeterioration KW - Fouling KW - Indicator PY - 2016 DO - https://doi.org/10.1080/08927014.2016.1177515 SN - 0892-7014 VL - 32 IS - 6 SP - 635 EP - 644 PB - Taylor & Francis Group CY - Abingdon AN - OPUS4-37337 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -