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Biology subjects

Bindschedler, S.

Publications and source records attributed to Bindschedler, S..

8 recordsLinked to original sources

Methods for evaluating bacterial dispersal on hyphal networks

In heterogeneous environments, the hyphae of filamentous fungi and oomycetes can facilitate the dispersal of other microorganisms. The use of these "fungal highways" (FH) is regulated by both physical and biological factors with their interplay resulting in variable capabilities of different microbes to establish FH. Several devices have been developed to test the movement of bacteria across mycelium. However, these methods are usually time-consuming and cannot be applied either at a large scale or in a high throughput format. In this study, we developed 3D-printed experimental devices that physically separate two environments while allowing hyphal networks to act as bridges for bacterial movement. The final design allows for the simultaneous testing of up to 10 pairs and the inclusion of any culturing media. With these devices, we investigated how fungal-bacterial pairing, nutrient conditions, and inoculation strategies influence FH formation. Bacterial transport was limited in nutrient-rich media but increased under poorer nutrient conditions, consistent with enhanced exploratory growth of the mycelium. Both cis- and trans-inoculation supported FH formation, although bacterial arrival was delayed in the absence of co-inoculation. The devices were used to demonstrate that transport of bacteria by FH was relevant for the colonization of a natural substrate. Finally, we established a novel in planta assay to evaluate FH formation during host colonization. This assay demonstrated that Fusarium graminearum can transport bacteria during wheat spike colonization. Together, these results provide accessible, scalable tools to study hyphal-mediated bacterial dispersal and highlight the combined role of biological specificity and nutrient context in the establishment of FH. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=75 SRC="FIGDIR/small/719220v1_ufig1.gif" ALT="Figure 1"> View larger version (25K): org.highwire.dtl.DTLVardef@1cf39aforg.highwire.dtl.DTLVardef@1d41e6corg.highwire.dtl.DTLVardef@1196637org.highwire.dtl.DTLVardef@85cc6c_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

A microfluidic platform for the observation and quantification of fungal highways

Soil is a complex system characterised by intra- and inter-kingdom interactions among microbial communities. While many different types of fungal-bacterial interactions have been described, hyphal-mediated transport of bacteria via the so-called "fungal highway" (FH) has not been mechanistically described. Bacteria require a liquid film for active movement; therefore, liquid saturation is a significant limiting factor for their dispersal in soil. Hyphal networks contribute to the connectivity between discrete soil microbial populations by providing a physical network through the unsaturated soil matrix. This network serves as a scaffold for liquid transport, thus allowing bacteria to migrate further or access previously isolated spaces. Studying these interactions is challenging due to the complex and stochastic nature of the soil environment; this "black box" aspect makes it difficult to visualise interactions at the microbial scale. Microfluidic technology can provide a solution by offering precise imaging at a high resolution in a physically and chemically controlled environment. We designed a microfluidic device, the Fungal Highways Device (FHD), that allows us to culture filamentous organisms in unsaturated environments and visualise and quantify bacterial dispersal along hyphal networks at the single-cell level. We showed that Pythium ultimum is essential for Pseudomonas putida movement across an unsaturated environment, and we identified mycelial biomass and hyphal front length as key factors influencing the bacterias movement towards the outlet. We propose that the liquid transport facilitated by P. ultimum mycelium influences the FH behaviour during its interaction with P. putida.

bioengineering↗

Co-isolation of Penicillium citrinum and its cell-switching partner Meyerozyma guilliermondii from a geothermal power plant

Progresses in geothermal energy and deep drilling technologies have opened a new window into the terrestrial subsurface. This provides direct access to deep geothermal fluids used to produce heat and electricity, creating an opportunity to isolate and characterize novel microbial strains from these extreme habitats. In this study, we report the co-isolation of two fungal strains. Penicillium citrinum (strain HEK1) was isolated first and thought to be axenic. However, upon exposure to stress (frost and ethanol), a second strain, corresponding to the dimorphic yeast Meyerozyma guilliermondii (strain HEK2), appeared in HEK1 cultures. Strain HEK2 appeared first in the cultures and was followed by the subsequent re-growth of strain HEK1, underscoring their close relationship. Moreover, strain HEK2, able to switch from yeast cells to pseudohyphae when growing alone, did not produce pseudohyphae when in direct contact with strain HEK1. Altogether, our results indicate an intricate interaction between these strains that may allow them to thrive in the deep subsurface. These two fungi represent the first fungal strains isolated from deep geothermal fluids. Their presence within the fluids was confirmed through molecular analysis. The isolation of these strains emphasizes the importance of considering fungi when investigating microbial diversity in subsurface geothermal environments. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=150 SRC="FIGDIR/small/599737v1_ufig1.gif" ALT="Figure 1"> View larger version (34K): org.highwire.dtl.DTLVardef@fdf699org.highwire.dtl.DTLVardef@148b751org.highwire.dtl.DTLVardef@fb37e9org.highwire.dtl.DTLVardef@113b911_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIFirst fungal strains isolated from a geothermal power plant C_LIO_LIThe two fungal strains were co-isolated from a geothermal fluid used for heat production C_LIO_LISurprising isolation of the cell-switching yeast upon stress exposure of an apparently axenic culture of the filamentous fungus C_LIO_LIFungal strains with high resistance to stressors and no apparent competition for carbon sources C_LI

microbiology↗

Microfluidic platform for microbial spore germination studies in multiple growth conditions

Spores are highly resistant dormant cells, adapted for survival and dispersal, that can withstand unfavourable environmental conditions for extended periods of time and later reactivate. Understanding the germination process of microbial spores is important in numerous areas including agriculture, food safety and health, and other sectors of biotechnology. Microfluidics combined with high-resolution microscopy allows to study spore germination at the single-cell level, revealing behaviours that would be hidden in standard population-level studies. Here, we present a microfluidic platform for germination studies where spores are confined to monolayers inside microchambers, allowing the testing of four growth conditions in parallel. This platform can be used with multiple species, including non-model organisms, and is compatible with existing image analysis software. In this study, we focused on three soil dwellers, two prokaryotes and one fungus, and revealed new insights into their germination. We studied endospores of the model bacterium Bacillus subtilis and demonstrated a correlation between spore density and germination in rich media. We then investigated the germination of the obligate-oxalotrophic environmental bacterium Ammoniphilus oxalaticus in a concentration gradient of potassium oxalate, showing that lower concentrations result in more spores germinating compared to higher concentrations. We also used this microfluidic platform to study the soil beneficial filamentous fungus Trichoderma rossicum, showing for the first time that the size of the spores and hyphae increase in response to increased nutrient availability, while germination times remain the same. Our platform allows to better understand microbial behaviour at the single-cell level, under a variety of controlled conditions. One-Sentence SummaryA microfluidic platform developed for spore germination studies in multiple growth conditions provides new insights into the germination of spores at the cellular level from three soil dwellers.

bioengineering↗

A ubiquitous Microcoleus species causes benthic cyanotoxic blooms worldwide

Recently, proliferations of benthic cyanobacteria producing derivatives of anatoxin-a have been reported in rivers all over the world. In three river systems, in New Zealand, the USA, and Canada, a cohesive cluster of Microcoleus strains was responsible for toxin production. Here, we document a similar toxigenic event that occurred at the mouth of the river Areuse in lake Neuchatel (Switzerland) and caused the death of several dogs. Using 16S RNA-based community analysis, we show that riverine benthic communities are dominated by Oscillatoriales and especially by Microcoleus strains. We correlate the detection of one sequence variant with the presence of anatoxin-a derivatives and use metagenomics to assemble a complete circular genome of the strain. The strain is distinct from the ones isolated in New Zealand, the USA, and Canada, but belongs to the same species; it shares significant traits with them, in particular a relatively small genome and incomplete vitamin biosynthetic pathways. Overall, our results suggest that the major anatoxin-a-associated benthic proliferations worldwide can be traced back to a single ubiquitous species, Microcoleus anatoxicus, rather than to a diversity of cyanobacterial lineages. We recommend that this species be monitored internationally in order to help predict and mitigate similar cyanotoxic events.

microbiology↗

Design and construction of 3D-printed devices to investigate active and passive bacterial dispersal on hydrated surfaces

To disperse in water-unsaturated environments, such as the soil, bacteria rely on the availability and structure of water films forming on biotic and abiotic surfaces, and, especially, along fungal mycelia. Dispersal along such "fungal highways" is driven both by mycelial physical properties and by interactions between bacteria and fungi. To understand the role of abiotic elements, we designed and 3D-printed two devices establishing stable liquid films that support bacteria dispersal in the absence of biotic interactions. The thickness of the liquid film determined the presence of hydraulic flow capable of carrying non-motile cells. In the absence of flow, only motile cells can disperse in the presence of an energy source. Non-motile cells could not disperse autonomously without flow, but dispersed when co-inoculated with motile cells. By teasing apart the abiotic and biotic dimensions, these 3D-printed devices will stimulate further research on microbial dispersal in soil and other water-unsaturated environments.

microbiology↗

Spatial scales of competition and a growth-motility tradeoff interact to determine bacterial coexistence

1.The coexistence of competing species is a long-lasting puzzle in evolutionary ecology research. Despite abundant experimental evidence showing that the opportunity for coexistence decreases as niche overlap increases between species, bacterial species and strains competing for the same resources are commonly found across diverse spatially heterogeneous habitats. We thus hypothesized that the spatial scale of competition may play a key role in determining bacterial coexistence, and interact with other mechanisms that promote coexistence, including a growth-motility tradeoff. To test this hypothesis, we let two Pseudomonas putida strains compete at local and regional scales by inoculating them either in a mixed droplet or in separate droplets in the same Petri dish, respectively. We also created conditions that allow the bacterial strains to disperse across abiotic or fungal hyphae networks. We found that competition at the local scale led to competitive exclusion while regional competition promoted coexistence. When competing in the presence of dispersal networks, the growth-motility tradeoff promoted coexistence only when the strains were inoculated in separate droplets. Our results provide a mechanism by which existing laboratory data suggesting competitive exclusion at a local scale is reconciled with the widespread coexistence of competing bacterial strains in complex natural environments with dispersal.

ecology↗

Exploiting oxalotrophic bacteria to control Aspergillus niger lung infections

Aspergillus fungi are opportunistic pathogens that affect a large number of people worldwide. Many aspects of Aspergillus spp. pathogenesis toward humans are known, but their ability to enhance their infectious potential by manipulating the environmental pH of its host has not been considered yet. In this study, we tested the hypothesis that by producing oxalic acid, Aspergillus niger can manipulate pH during lung infection and thus, interfering with this process could limit pathogenicity. To test this hypothesis, we co-cultured A. niger with oxalotrophic bacteria in increasingly complex testing systems (Petri dishes and 3D-cell cultures systems). In in vitro tests, oxalotrophic bacteria limit oxalic acid production and suppressed the pH shift induced by A. niger. In 3D-cell cultures (Transwells(R) and Bronchioles-on-a-chip), A. niger also modified pH, Ca2+ and oxalic acid concentrations. Co-inoculation with as little as 10 cells of the oxalatrophic bacterium strongly inhibited the germination and development of A. niger and returned each of the three parameters to the baseline physiological values of uninfected cells. This biocontrol interaction between oxalotrophic bacteria and oxalate-producing A. niger could represent a paradigm shift in the fight against opportunistic fungal pathogens, where the host environment is rendered less permissive to fungal development.

microbiology↗