Pseudomonas aeruginosa contaminations in tap water systems have caused severe health problems in both hospital and household settings. To ensure fast and reliable detection, culture-independent methods are recommendable. However, the typically low cell number in water samples requires sample enrichment prior to analysis. Therefore, we developed and optimized an adsorption elution method using monolithic adsorption filtration and subsequent centrifugal ultrafiltration that can be combined with culture-independent detection methods. The principle of adsorption of Pseudomonas aeruginosa by hydrophobic and ionic interactions was studied in modified epoxy-based monoliths. Optimized conditions (5-L initial sample volume at pH 3 filtered for 30 min through hydrolyzed monoliths (MAF-OH) and eluted with beef extract glycine buffer at pH 9.5) achieved a recovery of 67.1 ± 1.2% and a concentration factor of 103. For the first time, we therefore present a culture-independent approach for rapid enrichment and subsequent molecular biological quantification of P. aeruginosa by qPCR from tap water samples by monolithic adsorption filtration. The total enrichment and quantification process takes 4 h. This work further stresses the versatility of the monolithic adsorption filtration and its possibilities as a concentration tool for culture-independent analytics of pathogenic bacteria in the environment.
Graphical abstract
Immer wieder kommt es zu bakteriellen Verunreinigungen in Trinkwasserleitungen. Diese können eine Gesundheitsgefahr darstellen und führen sowohl zu großer Besorgnis in der Öffentlichkeit als auch zu wirtschaftlichen Problemen, wenn die Ursache nicht zeitnah beseitigt wird. Während Escherichia coli und Enterococcus faecalis primär durch fäkale Verunreinigungen in Trinkwassersysteme gelangen können, kann Pseudomonas aeruginosa als Boden‐ und Wasserkeim in Leitungs‐, Mineral‐, Tafel‐ oder Schwimmbadwasser vorkommen [1]. P. aeruginosa kann sich auf nahezu allen feuchten Oberflächen ansiedeln, überlebt auch unter extremen Bedingungen (40°C, Trinkwasser mit sehr geringer Nährstoffdichte, niedrige pH‐Werte), bildet Biofilme und ist als nosokomialer Keim für viele krankenhausassoziierte Infektionen verantwortlich. Als sogenannter opportunistischer pathogener Keim ist P. aeruginosa besonders für immungeschwächte Personen gefährlich. Da P. aeruginosa intrinsisch bereits gegen eine Vielzahl an Antibiotika resistent ist und per horizontalem Gentransfer weitere Resistenzen erwerben kann, wird die Behandlung einer P. aeruginosa‐Infektion zunehmend herausfordernder. Dementsprechend sollten Verunreinigungen von Trinkwasserleitungen mit P. aeruginosa möglichst frühzeitig erkannt und bekämpft werden.
<p>Antimicrobials (AM) play a critical role in the treatment of human and animal (aquatic and terrestrial) diseases, which has led to their widespread application and use. Antimicrobial resistance (AMR) is the ability of microorganisms (e.g. bacteria, viruses and some parasites) to stop an antibiotic, such as an antimicrobial, antiviral or antimalarial, from working against them. Globally, about 700 000 deaths per year arise from resistant infections as a result of the fact that antimicrobial drugs have become less effective at killing resistant pathogens. Antimicrobial chemicals that are present in environmental compartments can trigger the development of AMR. These chemicals can also cause antibiotic-resistant bacteria (ARB) to further spread antibiotic resistance genes (ARG) because they may have an evolutionary advantage over non-resistant bacteria. Thus, AMR is a global threat to health, livelihoods and the achievement of the Sustainable Development Goals, both in developing and developed countries. For some time now, antimicrobial resistance (AMR) has been approached mainly from the human and animal health angles, however little is known about the impacts that AMR in the environment may have on health. A better understanding of how antimicrobial resistance moves from agricultural areas to the environment through soil and water is important if we are to develop guidance to managing it cost effectively. We examined the potential of nuclear techniques&#8212;the application of compound-specific stable isotope analysis (CSIA)&#8212;as a powerful tool to determine the source and fate of antibiotics in the environment and detect the degradation of antibiotics by transformation-induced isotopic effects. CSIA can be used to qualify and quantify in situ transformations. The latest methodological advances even allow the analysis of several elements (H, C, Cl, N) within a molecule This multi-element isotope information is used to elucidate in-situ transformation pathways and underlying reaction mechanisms.</p>
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