Evaluation of environmental scanning electron microscopy for analysis of Proteus mirabilis crystalline biofilms in situ on urinary catheters

Proteus mirabilis is a common cause of catheter-associated urinary tract infections and frequently leads to blockage of catheters due to crystalline biofilm formation. Scanning electron microscopy (SEM) has proven to be a valuable tool in the study of these unusual biofilms, but entails laborious sample preparation that can introduce artefacts, undermining the investigation of biofilm development. In contrast, environmental scanning electron microscopy (ESEM) permits imaging of unprocessed, fully hydrated samples, which may provide much insight into the development of P. mirabilis biofilms. Here, we evaluate the utility of ESEM for the study of P. mirabilis crystalline biofilms in situ, on urinary catheters. In doing so, we compare this to commonly used conventional SEM approaches for sample preparation and imaging. Overall, ESEM provided excellent resolution of biofilms formed on urinary catheters and revealed structures not observed in standard SEM imaging or previously described in other studies of these biofilms. In addition, we show that energy-dispersive X-ray spectroscopy (EDS) may be employed in conjunction with ESEM to provide information regarding the elemental composition of crystalline structures and demonstrate the potential for ESEM in combination with EDS to constitute a useful tool in exploring the mechanisms underpinning crystalline biofilm formation

Worldwide tests of generic attractants, a promising tool for early detection of non-native cerambycid species

A large proportion of the insects which have invaded new regions and countries are emerging species, being found for the first time outside their native range. Being able to detect such species upon arrival at ports of entry before they establish in non-native countries is an urgent challenge. The deployment of traps baited with broad-spectrum semiochemical lures at ports-of-entry and other high-risk sites could be one such early detection tool. Rapid progress in the identification of semiochemicals for cerambycid beetles during the last 15 years has revealed that aggregation-sex pheromones and sex pheromones are often conserved at global levels for genera, tribes or subfamilies of the Cerambycidae. This possibly allows the development of generic attractants which attract multiple species simultaneously, especially when such pheromones are combined into blends. Here, we present the results of a worldwide field trial programme conducted during 2018-2021, using traps baited with a standardised 8-pheromone blend, usually com-plemented with plant volatiles. A total of 1308 traps were deployed at 302 sites covering simultaneously or sequentially 13 European countries, 10 Chinese provinces and some regions of the USA, Canada, Australia, Russia (Siberia) and the Caribbean (Martinique). We intended to test the following hypotheses: 1) if a species is regularly trapped in significant numbers by the blend on a continent, it increases the prob-ability that it can be detected when it arrives in other countries/continents and 2) if the blend exerts an effective, generic attraction to multiple species, it is likely that previously unknown and unexpected spe-cies can be captured due to the high degree of conservation of pheromone structures within related taxa. A total of 78,321 longhorned beetles were trapped, representing 376 species from eight subfamilies, with 84 species captured in numbers greater than 50 individuals. Captures comprised 60 tribes, with 10 tribes including more than nine species trapped on different continents. Some invasive species were captured in both the native and invaded continents. This demonstrates the potential of multipheromone lures as ef-fective tools for the detection of 'unexpected' cerambycid invaders, accidentally translocated outside their native ranges. Adding new pheromones with analogous well-conserved motifs is discussed, as well as the limitations of using such blends, especially for some cerambycid taxa which may be more attracted by the trap colour or other characteristics rather than to the chemical blend

Regelgeleitete Konstruktion von statistischen Textaufgaben. Anwendung von linear logistischen Testmodellen und Aufgabencloning. Projekt Regelgeleitete Itementwicklung

Im vorliegenden Projekt werden Inhalte der kognitiven Psychologie, Psychometrie und Computerwissenschaften kombiniert, um einen automatischen Aufgabengenerator zu entwickeln, der unter Anwendung von linear-logistischen Testmodellen, optimalem Design und Modellen für Aufgabencloning adaptiv individuell maßgeschneiderte statistische Textaufgaben produziert. Mit Hilfe dieser Testkonstruktionsprinzipien wird effiziente Kompetenzdiagnostik im mathematisch-statistischen Bereich für Oberstufenschüler und Studenten ermöglicht. Die bisherigen Projektergebnisse sind vielversprechend: Verschiedenste kognitive Komponenten konnten als signifikante schwierigkeitsgenerierende Merkmale identifiziert und in halbautomatischer Aufgabenkonstruktion als Vorstufe zur vollautomatischen Generierung mit Hilfe von Textbausteinen umgesetzt werden (beispielhaft wird ein Test zu Konfidenzintervallen dargestellt). Die Ergebnisse zeigen die Anwendbarkeit von regelgeleiteter und automatischer Aufgabengenerierung auf Textaufgaben mit Statistikinhalten und dienen nun zur Verfeinerung des automatischen Aufgabengenerators. Die konstruierten Aufgaben haben einen guten Rasch-Fit und stellen den Ausgangspunkt für verfeinerte inhaltliche und konstruktionstechnische Weiterentwicklungen und adaptive Implementierungen dar. Verschiedene mögliche Modelle für die Kalibrierung der automatisch generierten Aufgaben werden aufgezeigt und ein Prototyp eines automatischen Aufgabengenerators für statistische Textaufgaben wird dargestellt. Außerdem wird eine Optimierung der statistischen Modelle und Prozeduren in Hinblick auf computergestützte adaptive Testungen, Kompetenzdiagnostik und groß angelegte Testdurchläufe verfolgt. (DIPF/Autor

Elucidating the genetic basis of crystalline biofilm formation in Proteus mirabilis

Proteus mirabilis forms extensive crystalline biofilms on urethral catheters that occlude urine flow and frequently complicate the management of long-term-catheterized patients. Here, using random transposon mutagenesis in conjunction with in vitro models of the catheterized urinary tract, we elucidate the mechanisms underpinning the formation of crystalline biofilms by P. mirabilis. Mutants identified as defective in blockage of urethral catheters had disruptions in genes involved in nitrogen metabolism and efflux systems but were unaffected in general growth, survival in bladder model systems, or the ability to elevate urinary pH. Imaging of biofilms directly on catheter surfaces, along with quantification of levels of encrustation and biomass, confirmed that the mutants were attenuated specifically in the ability to form crystalline biofilms compared with that of the wild type. However, the biofilm-deficient phenotype of these mutants was not due to deficiencies in attachment to catheter biomaterials, and defects in later stages of biofilm development were indicated. For one blocking-deficient mutant, the disrupted gene (encoding a putative multidrug efflux pump) was also found to be associated with susceptibility to fosfomycin, and loss of this system or general inhibition of efflux pumps increased sensitivity to this antibiotic. Furthermore, homologues of this system were found to be widely distributed among other common pathogens of the catheterized urinary tract. Overall, our findings provide fundamental new insight into crystalline biofilm formation by P. mirabilis, including the link between biofilm formation and antibiotic resistance in this organism, and indicate a potential role for efflux pump inhibitors in the treatment or prevention of P. mirabilis crystalline biofilms