Are anaerobic fungi crucial hidden players of microbiomes in anoxic environment?

Anaerobic fungi are known to migrate and establish a 3D network of biofilms (microbiomes) and live invisible in the rumen and terrestrial subsurface, deep-sea - marine, and anoxic environment. They deserve our attention to understand anoxic fungal ecology and functions and develop new products and solutions. Such fungi activate unique genes to produce various polysaccharidases deemed essential for degrading plants' lignocellulosic materials. Nutrient release, recycling, and physical support by anaerobic fungi are crucial for microbiome formation. Multiple reports point to the ability of strictly anaerobic and facultative fungi to adapt and live in anoxic subsurface. Deep-sea sediments and natural anoxic methane-emitting salty waters of sulfidic springs offer suitable habitats for developing prokaryotic-fungal microbiomes. Researchers found a billion-year-old fossil of the fungus-prokaryotic sulfate-reducing consortium buried in deep-sea biospheres. Fungal spores' ability to migrate, even after germination, through sandy layers demonstrates their potential to move up and down porous geological layers or rock fissures. Selective fungal affinity to specific wood in wood chip arrays might help differentiate viable anaerobic fungi from an anoxic environment for their rapid collection and investigation. New collection methods, cultivation, gene expression, and drug and enzyme activity analyses can boost anaerobic fungal research.

Facilitating hand hygiene in displacement camps during the COVID-19 pandemic: a qualitative assessment of a novel handwashing stand and hygiene promotion package.

BACKGROUND: Handwashing with soap is critical for the prevention of diarrhoeal diseases and outbreak related diseases, including interrupting the transmission of COVID-19. People living in large displacement settings are particularly vulnerable to such outbreaks, however, practicing handwashing is typically challenging in these contexts. METHODS: We conducted a qualitative assessment of the implementation of a combined intervention to facilitate handwashing behaviour in displacement camps and in surrounding communities in Bangladesh, Ethiopia and the Democratic Republic of Congo during the COVID-19 pandemic. The intervention comprised a 'hardware' infrastructural component (provision of the Oxfam Handwashing Station) and a 'software' hygiene promotion package (Mum's Magic Hands). We used programmatic logbooks, interviews with implementation staff and focus group discussions with crisis-affected populations to assess the use, feasibility and acceptability of the intervention. RESULTS: Both components of the intervention were viewed as novel and appealing by implementing staff and crisis-affected populations across the study sites. The acceptability of the handwashing station could be improved by redesigning the tap and legs, exploring local supply chain options, and by providing a greater number of facilities. The implementation of the hygiene promotion package varied substantially by country making it challenging to evaluate and compare. A greater focus on community engagement could address misconceptions, barriers related to the intuitiveness of the handwashing station design, and willingness to participate in the hygiene promotion component. CONCLUSIONS: The combination of a 'hardware' and 'software' intervention in these settings appeared to facilitate both access and use of handwashing facilities. The acceptability of the combined intervention was partially because a great deal of effort had been put into their design. However, even when delivering well-designed interventions, there are many contextual aspects that need to be considered, as well as unintended consequences which can affect the acceptability of an intervention.

Designing and Evaluating Multimodal Interactions for Facilitating Visual Analysis With Dashboards.

Exploring and analyzing data using visualizations is at the heart of many decision-making tasks. Typically, people perform visual data analysis using mouse and touch interactions. While such interactions are often easy to use, they can be inadequate for users to express complex information and may require many steps to complete a task. Recently natural language interaction has emerged as a promising technique for supporting exploration with visualization, as the user can express a complex analytical question more easily. In this paper, we investigate how to synergistically combine language and mouse-based direct manipulations so that the weakness of one modality can be complemented by the other. To this end, we have developed a novel system, named Multimodal Interactions System for Visual Analysis (MIVA), that allows user to provide input using both natural language (e.g., through speech) and direct manipulation (e.g., through mouse or touch) and presents the answer accordingly. To answer the current question in the context of past interactions, the system incorporates previous utterances and direct manipulations made by the user within a finite-state model. The uniqueness of our approach is that unlike most previous approaches which typically support multimodal interactions with a single visualization, MIVA enables multimodal interactions with multiple coordinated visualizations of a dashboard that visually summarizes a dataset. We tested MIVA's applicability on several dashboards including a COVID-19 dashboard that visualizes coronavirus cases around the globe. We further empirically evaluated our system through a user study with twenty participants. The results of our study revealed that MIVA system enhances the flow of visual analysis by enabling fluid, iterative exploration and refinement of data in a dashboard with multiple-coordinated views.

Searching the Visual Style and Structure of D3 Visualizations.

We present a search engine for D3 visualizations that allows queries based on their visual style and underlying structure. To build the engine we crawl a collection of 7860 D3 visualizations from the Web and deconstruct each one to recover its data, its data-encoding marks and the encodings describing how the data is mapped to visual attributes of the marks. We also extract axes and other non-data-encoding attributes of marks (e.g., typeface, background color). Our search engine indexes this style and structure information as well as metadata about the webpage containing the chart. We show how visualization developers can search the collection to find visualizations that exhibit specific design characteristics and thereby explore the space of possible designs. We also demonstrate how researchers can use the search engine to identify commonly used visual design patterns and we perform such a demographic design analysis across our collection of D3 charts. A user study reveals that visualization developers found our style and structure based search engine to be significantly more useful and satisfying for finding different designs of D3 charts, than a baseline search engine that only allows keyword search over the webpage containing a chart.

Multimetal bioremediation and biomining by a combination of new aquatic strains of Mucor hiemalis.

Here we describe a unique microbial biotechnology for simultaneous bioremediation and biomining of twelve ionic metals overcoming the obstacles of multimetal toxicity to microbes. After a thorough search of key microorganisms in microbiomes of many sulfidic springs in Bavaria (Germany) over an area of 200 km2, we found three new strains EH8, EH10 and EH11 of Mucor hiemalis physiologically compatible and capable of multimetal-remediation and enrichment. We combined the multimetal-resistance, hyper-accumulation and elicitation power of EH8, EH10 and EH11 to develop a novel biotechnology for simultaneous removal, fractionation and enrichment of metal ions. As a first step we showed the intracellular fixing and deposition of mercury as nanospheres in EH8's sporangiospores. Scanning Electron Microscopy-Energy-Dispersive X-Ray analysis revealed binding and precipitation of other applied metal ions as spherical nano-particles (~50-100 nm) at the outer electro-negative cellwall-surface of EH8, EH10 and EH11 sporangiospores. Microbiomes, germinated spores and dead insoluble cellwalls of these strains removed >81-99% of applied Al, Cd, Co, Cr, Cu, Hg, Ni, Pb, U, and Zn simultaneously and furthermore enriched precious Ag, Au and Ti from water all within 48 h, demonstrating the potential of new biotechnologies for safe-guarding our environment from metal pollution and concentrating precious diluted, ionic metals.

Detection of the formyl radical by EPR spin-trapping and mass spectrometry.

For the first time we here present the unambiguous identification of the formyl radical (•CHO) by EPR (Electron Paramagnetic Resonance) spectroscopy and mass spectrometry (MS) using DMPO (5,5-dimethyl-1-pyrroline N-oxide) as spin trap at ambient temperature without using any catalyst(s). The •CHO was continuously generated by UV photolysis in closed anoxic environment from pure formaldehyde (HCHO) in aqueous solution. The isotropic hyperfine structure constants of •CHO were determined as aN = 15.72G and aH = 21.27G. The signals were deconvoluted and split by simulation in their single adduct components: DMPO-CHO, DMPO-H and DMPO-OH. We verified our results at first using MNP (2-methyl-2-nitroso-propane) as spin trap with known literature data and then mass spectrometry. Similarly the MNP adduct components MNP-CHO, MNP-H as well as its own adduct, the MNP-2-methyl-2-propyl (MNP-MP) were deconvoluted. Due to the low signal intensities, we had to accumulate single measurements for both spin traps. Using MS we got the exact mass of the reduced •CHO adduct independently confirming the result of EPR detection of formyl radical.

Applying Pragmatics Principles for Interaction with Visual Analytics.

Interactive visual data analysis is most productive when users can focus on answering the questions they have about their data, rather than focusing on how to operate the interface to the analysis tool. One viable approach to engaging users in interactive conversations with their data is a natural language interface to visualizations. These interfaces have the potential to be both more expressive and more accessible than other interaction paradigms. We explore how principles from language pragmatics can be applied to the flow of visual analytical conversations, using natural language as an input modality. We evaluate the effectiveness of pragmatics support in our system Evizeon, and present design considerations for conversation interfaces to visual analytics tools.

Responses of the antioxidative and biotransformation enzymes in the aquatic fungus Mucor hiemalis exposed to cyanotoxins.

OBJECTIVES: To investigate antioxidative and biotransformation enzyme responses in Mucor hiemalis towards cyanotoxins considering its use in mycoremediation applications. RESULTS: Catalase (CAT), glutathione reductase (GR), and glutathione peroxidase (GPx) in M. hiemalis maintained their activities at all tested microcystin-LR (MC-LR) exposure concentrations. Cytosolic glutathione S-transferase (GST) activity decreased with exposure to 100 µg MC-LR l-1 while microsomal GST remained constant. Cylindrospermopsin (CYN) at 100 µg l-1 led to an increase in CAT activity and inhibition of GR, as well as to a concentration-dependent GPx inhibition. Microsomal GST was inhibited at all concentrations tested. ß-N-methylamino-L-alanine (BMAA) inhibited GR activity in a concentration-dependent manner, however, CAT, GPx, and GST remained unaffected. CONCLUSIONS: M. hiemalis showed enhanced oxidative stress tolerance and intact biotransformation enzyme activity towards MC-LR and BMAA in comparison to CYN, confirming its applicability in bioreactor technology in terms of viability and survival in their presence.

Ecology, adaptation, and function of methane-sulfidic spring water biofilm microorganisms, including a strain of anaerobic fungus Mucor hiemalis.

Ecological aspects, adaptation, and some functions of a special biofilm and its unique key anaerobic fungus Mucor hiemalis strain EH11 isolated from a pristine spring (Künzing, Bavaria, Germany) are described. The spring's pure nature is characterized by, for example, bubbling methane, marine-salinity, mild hydrothermal (~19.1°C), sulfidic, and reductive-anoxic (Eh : -241 to -253 mV, O2 : ≤ 0.1 mg/L) conditions. It is geoecologically located at the border zone between Bavarian Forest (crystalline rocky mountains) and the moor-like Danube River valley, where geological displacements bring the spring's water from the deeper layers of former marine sources up to the surface. In the spring's outflow, a special biofilm with selective microorganisms consisting of archaea, bacteria, protozoa (ciliate), and fungus was found. Typical sulfidic-spring bryophyta and macrozoobenthos were missing, but many halo- and anaerotolerant diatoms and ciliate Vorticella microstoma beside EH11 were identified. Phase contrast and scanning electron microscopy revealed the existence of a stabilizing matrix in the biofilm formed by the sessile fungal hyphae and the exopolysaccharide substance (EPS) structures, which harbors other microorganisms. In response to ecological adaptation pressure caused by methane bubbles, EH11 developed an atypical spring-like hyphal morphology, similar to the spiral stalk of ciliate V. microstoma, to rise up with methane bubbles. For the first time, it was also demonstrated that under strict anaerobic conditions EH11 changes its asexual reproduction process by forming pseudosporangia via hyphal cell divisions as well as switching its metabolism to chemoautotrophic bacteria-like anaerobic life using acetate as an e-donor and ferrihydrite as an e-acceptor, all without fermentation. EH11 can be suggested to be useful for the microbial community in the Künzing biofilm not only due to its physical stabilization of the biofilm's matrix but also due to its ecological functions in element recycling as well as a remover of toxic metals.

A new mercury-accumulating Mucor hiemalis strain EH8 from cold sulfidic spring water biofilms.

Here, we report about a unique aquatic fungus Mucor hiemalisEH8 that can remove toxic ionic mercury from water by intracellular accumulation and reduction into elemental mercury (Hg0 ). EH8 was isolated from a microbial biofilm grown in sulfidic-reducing spring water sourced at a Marching's site located downhill from hop cultivation areas with a history of mercury use. A thorough biodiversity survey and mercury-removal function analyses were undertaken in an area of about 200 km2 in Bavaria (Germany) to find the key biofilm and microbe for mercury removal. After a systematic search using metal removal assays we identified Marching spring's biofilm out of 18 different sulfidic springs' biofilms as the only one that was capable of removing ionic Hg from water. EH8 was selected, due to its molecular biological identification as the key microorganism of this biofilm with the capability of mercury removal, and cultivated as a pure culture on solid and in liquid media to produce germinating sporangiospores. They removed 99% of mercury from water within 10-48 h after initial exposure to Hg(II). Scanning electron microscopy demonstrated occurrence of intracellular mercury in germinating sporangiospores exposed to mercury. Not only associated with intracellular components, but mercury was also found to be released and deposited as metallic-shiny nanospheres. Electron-dispersive x-ray analysis of such a nanosphere confirmed presence of mercury by the HgMα peak at 2.195 keV. Thus, a first aquatic eukaryotic microbe has been found that is able to grow even at low temperature under sulfur-reducing conditions with promising performance in mercury removal to safeguard our environment from mercury pollution.

Induction of glutathione S-transferase in biofilms and germinating spores of Mucor hiemalis strain EH5 from cold sulfidic spring waters.

The occurrence and activation of glutathione S-transferase (GST) and the GST activities in biofilms in cold sulfidic spring waters were compared to the occurrence and activation of GST and the GST activities of the aquatic fungal strains EH5 and EH7 of Mucor hiemalis isolated for the first time from such waters. Using fluorescently labeled polyclonal anti-GST antibodies and GST activity measurements, we demonstrated that a high level of GST occurred in situ in natural biofilms and pure cultures of strain EH5. Measurement of microsomal and cytosolic soluble GST activities using different xenobiotic substrates, including 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene, 1,2-epoxy-3-(4-nitrophenoxy)propane, 1-iodo-2,4-dinitrobenzene, and fluorodifen, showed that the overall biotransforming abilities of biofilms were at least sixfold greater than that of strain EH5 alone. Increasing the level of sodium thiosulfate (STS) in the medium stimulated the microsomal and cytosolic GST activities with CDNB of strain EH5 about 44- and 94-fold, respectively, compared to the activities in the control. The induction of microsomal GST activity with fluorodifen by STS was strongly linear, but the initial strong linear increase in cytosolic GST activity with fluorodifen showed saturation-like effects at STS concentrations higher than approximately 1 mM. Using laser scanning confocal and conventional fluorescence microscopy, abundant fluorescently labeled GST proteins were identified in germinating sporangiospores of strain EH5 after activation by STS. High-performance size exclusion chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of at least two main GSTs ( approximately 27.8- and approximately 25.6-kDa subunits) in the cytosol of EH5, whereas the major 27.8-kDa subunit was the only GST in microsomes. We suggest that differential cellular GST expression takes place in strain EH5 depending on spore and hyphal development. Our results may contribute to our understanding of induction of GST by sulfurous compounds, as well as to the immunofluorescence visualization of GST in aquatic fungus and fungus-bacterium biofilms.

Meningoencephalitis as a presentation of Kawasaki disease.

We describe an 11-year-old boy who presented with meningoencephalitis and was found later to have Kawasaki disease. The neurologic outcome of this patient was excellent, with full recovery without any neurologic deficit. The case report is followed by a review of the neurologic associations and complications of Kawasaki disease.

Influence of ionic strength and organic modifier concentrations on characterization of aquatic fulvic and humic acids by high-performance size-exclusion chromatography.

Aquatic fulvic acid (FA) and humic acid (HA) were characterized by an aqueous high-performance size-exclusion chromatography (HPSEC) using a hydrophilic polymeric stationary phase and an aqueous eluent at neutral pH and low-ionic strength (5 mM Na2HPO4; final ionic strength, 13 mM). Employed HPSEC showed low sensitivity of FA to variations in ionic strength (13 and 100 mM) and contents of organic modifier (0.1 or 40% methanol) in aqueous eluent. Under these analytical conditions, peak maxima of relative UV signals versus molecular mass (Mr) defined as M'p and peak maxima of relative mass concentrations versus Mr defined as Mp of FA were shown to be located at 548-690 and 500, respectively. Organic modifier concentrations of 40% methanol in aqueous eluent enabled not only analysis of FA, but also analysis of some aquatic HA by HPSEC. Analysis showed M'p and Mp values of aquatic HAs around 1000 and 600, respectively. Measured molecular mass data of FA were found to be consistent with the recently published data describing low molecular masses of FA. Results recommend the use of the described HPSEC as a simple, rapid, reproducible, low-cost method giving consistent molecular sizes/masses of FA and some aquatic HAs.

Uptake, metabolism, and effects on detoxication enzymes of isoproturon in spawn and tadpoles of amphibians.

Using ring-(14)C-labeled isoproturon (1 microg/L), the uptake into spawn and tadpoles of Bombina bombina and Bombina variegata was investigated. Two percent of the applied radioactivity was found per gram fresh weight in the embryo after 24h. Results indicate that the jelly mass of the spawn does not act as a sufficient physical barrier for protection against the uptake and influence of isoproturon (IPU) on the embryo. In vivo metabolism of ring-14C-labeled IPU by the cytochrome P-450 system was analyzed in tadpoles. Different metabolites of IPU, such as N-demethylated and C-hydroxylated derivatives, and the olefinic metabolite were detected. In tadpoles of B. variegata, the activity of microsomal and soluble glutathione-S-transferase (sGSTs) toward different model substrates was measured after treatment with IPU. Activities of sGST increased corresponding to elevated stress by IPU dependent on exposure time and dose. Compared to the pure active ingredient IPU, the commercial phenyl-urea herbicide Tolkan Flo, consisting of IPU and an emulsifier, also caused significantly elevated enzymatic response.