1

EQUIPMENT

2

RESEARCH

3

Service

News

For the following Paper we used the high throughput phenotyping capability of the BiMM equipment to screen more than 1300 clinical Candida isolates for drug sensitivity.
With robotic help Reinhard Beyer produced 1400 96-well plates with a concentration gradient of eight antifungals and generated and analyzed 140 000 growth curves.

16.01.2019

Antifungal susceptibility of yeast blood stream isolates collected during a 10 year period in Austria.

Beyer R, Spettel K, Zeller I, Lass-Flörl C, Achleitner D, Krause R, Apfalter P, Buzina W, Strauss J, Gregori C, Schüller C, Willinger B.

Mycoses 2019 Jan 12. doi: 10.1111/myc.12892. [Epub ahead of print]
PMID: 30636016

https://www.ncbi.nlm.nih.gov/pubmed/30636016

BACKGROUND:
Candida-associated infections put a significant burden on western health care systems. Development of (multi-) resistant fungi can become untreatable and threaten especially vulnerable target groups, such as the immunocompromised.
OBJECTIVES:
We assessed antifungal susceptibility and explored possible influence factors of clinical Candida isolates collected from Austrian hospitals between 2007 and 2016 METHODS: 1360 clinical Candida spp. isolated from blood cultures were subjected to antifungal susceptibility testing (AFST) in a liquid handling aided continuous microdilution assay. We tested against fluconazole, voriconazole, posaconazole, itraconazole, isavuconazole, anidulafungin, caspofungin and micafungin according to EUCAST with additional recording of growth curves. We performed rigid quality control on each assay via growth curve assessment and included two standard reference strains. Minimal inhibitory concentrations (MIC) were quantified according to EUCAST guideline E.DEF 7.3.1 and susceptibility was evaluated using EUCAST clinical breakpoints.
RESULTS:
The isolate collection consisted of C. albicans (59%), C. glabrata (19%), C. parapsilosis (9%), C. tropicalis (5%) and C. krusei (3%) and few other Candida species and fungi (5%). During the observed time period, species abundance as well as antifungal resistance rates remained constant. Multi-resistance was rare and we found no single isolate which was resistant to both azoles and echinocandins. Within the antifungal resistance profile of our strain collection, we observed clusters along species boundaries.
CONCLUSIONS:
Over the last decade, the distribution of Candida species and its level of antifungal resistance remained constant in Austria. Our data compares well with other European countries. Principal component analysis of the susceptibility profile of this collection revealed species-specific clusters and substantial intra-species variation, especially for C. glabrata.

We proudly present our new paper with contributions of the BiMM facility.

04.09.2018

Competition of Candida glabrata against Lactobacillus is Hog1 dependent.

Beyer R, Jandric Z, Zutz C, Gregori C, Willinger B, Jacobsen ID, Kovarik P, Strauss J, Schüller C.

Cell Microbiol. 2018 Aug 15:e12943. doi: 10.1111/cmi.12943. [Epub ahead of print]
PMID: 30112857

Candida glabrata is a common human fungal commensal and opportunistic pathogen. This fungus shows remarkable resilience as it can form recalcitrant biofilms on indwelling catheters, has intrinsic resistance against azole antifungals and is causing vulvo-vaginal candidiasis. As a nosocomial pathogen, it can cause life-threatening bloodstream infections in immune-compromised patients. Here, we investigate the potential role of the high osmolarity glycerol response (HOG) MAP kinase pathway for C. glabrata virulence. The C. glabrata MAP kinase CgHog1 becomes activated by a variety of environmental stress conditions such as osmotic stress, low pH and carboxylic acids and subsequently accumulates in the nucleus. We found that CgHog1 allows C. glabrata to persist within murine macrophages but it is not required for systemic infection in a mouse model. C. glabrata and Lactobacilli co-colonize mucosal surfaces. Lactic acid at a concentration produced by vaginal Lactobacillus spp. causes CgHog1 phosphorylation and accumulation in the nucleus. In addition, CgHog1 enables C. glabrata to tolerate different Lactobacillus spp. and their metabolites when grown in co-culture. Using a phenotypic diverse set of clinical C. glabrata isolates, we find that the HOG pathway is likely the main quantitative determinant of lactic acid stress resistance. Taken together, our data indicate that CgHog1 has an important role in the confrontation of C. glabrata with the common vaginal flora.

Articel describing new species from Donau and two new combinations has been accepted for publication in journal Nova Hedwigia

Mai 2018

„Metapochonia lutea, a new species isolated from the Danube river in Austria“

A new species Metapochonia lutea (Ascomycota, Hypocreales) is described and illustrated. This fungus differs from the other taxa in the genus Metapochonia by its production of intensive yellow pigment in culture, conidiophores with relatively complex verticillate branching, bean-shaped conidia, and by delayed development of one-celled and prominent thick-walled submerged chlamydospores usually in chains or irregular clusters of 3-5 cells. The new taxon is well supported by phylogenetic analysis of the internal transcribed spacer region (ITS) and translation elongation factor-1α gene (TEF-1α). Two new combinations are proposed for Pochonia parasitica and Pochonia cordycepisociata, as both species are resolved in a phylogenetic clade with other Metapochonia species.

Roman Labuda (1)*, Andreas Bernreiter (2), Christoph Schüller (1), Alena Kubátová (3), Roland Hellinger (4) & Joseph Strauss (2)

1 University of Veterinary Medicine (VetMed), Bioactive Microbial Metabolites group (BiMM), University and Research Center (UFT) – Campus Tulln,
Konrad Lorenz Strasse 24, 3430 Tulln a.d. Donau, Austria
*Corresponding author. Email: roman.labuda@vetmeduni.ac.at
2 Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Vienna (BOKU); Konrad Lorenz Strasse 24, 3430 Tulln a.d. Donau, Austria
3 Charles University, Faculty of Science, Department of Botany, Culture Collection of Fungi (CCF),
Benátská 2, 128 01 Prague 2, Czech Republic
4 Department of Agrobiotechnology, Center for Analytical Chemistry, University of Natural Resources and Life Sciences, Vienna (BOKU); Konrad Lorenz Strasse 20, 3430 Tulln a.d. Donau, Austria

https://www.schweizerbart.de/journals/nova_hedwigia

Flash Chromatography Machine installed at BiMM

March 2018

puriFlash 450 system

A State of the Art machine puriFlash 450 is in place from now on for our
large number of purifications.

http://www.interchim.com/pp.php?ref=95

Talk and poster at Final Scientific Conference of ImResFun

09.2017

The BiMM robots alow efficient high throughput analysis of growth phenotypes. We investigated growth under diverse conditions of 240 C. glabrata strains originating from the Vienna General Hospital. The results were presented at the Final Scientific Conference of ImResFun.

Talk and poster with the title:
The broad and malleable phenotypic space of clinical Candida glabrata strains. A high throughput mapping of phenotypes reveals highly individualistic adaptations to a fluctuating environment.

Candida glabrata is a successful commensal and able to adapt to a variety of environmental situations and a cause of vulvo-vaginal candidiasis and shows high resistance to azole antifungals. To explore how this flexibility is possible, we investigated the phenotypic variation of several hundred C.glabrata clinical isolates collected by the Vienna General Hospital. Our quantitative high-throughput determination of growth parameters revealed a broad variation of phenotypes. For example temperature influenced growth rate in an unexpected range. We found that many strains tolerated either higher or lower temperatures while all strains had a temperature optimum at around 39°C. The collection was also scored for resistance to low pH, weak acid, osmotic stress, carbon source utilization and resistance to antifungals. Interestingly, the population spread for heat stress resistance while it remained confined for osmitic stress. This argues for the polygenetic basis of heat stress versus only few key processes contribute to osmotic stress.

Furthermore, about 15% of these strains are efficiently forming biofilms. We further investigated the stability of some phenotypes. We tried
to manipulate fluconazole resistance by adapting sensitive strains to intermediate fluconazole concentrations. This succeeded but lead to a simultaneous post-adaption loss of growth rate on rich medium, indicating that adaptation to fluconazole comes at a cost. Genome sequencing of selected phenotypically eye-catching strains revealed that their genomic profiles differed in up to 105 bases making specific assignments difficult.

For stress resistance we analyzed the general role of CgHog1 and found a widely differing spectrum of basal Hog1 phosphorylation and stress response rates. The overall change of CgHog1 phosphorylation correlated with stress resistance of the particular strains. We conclude that these C. glabrata clinical isolates exhibit a broad phenotypic spectrum. Further investigation of the genetic and epigenetic basis of these variations and their stability are required to understand the success of C. glabrata as an opportunistic commensal.

contributing authors:
Christoph Schüller, Reinhard Beyer, Birgit Willinger, Joseph Strauss
1 Universität für Bodenkultur, Konrad Lorenz Strasse 24, Tulln a.d.
Donau 3430, Austria
2 Medizinische Universität Wien, Klinische Abteilung für Klinische
Mikrobiologie Währinger Gürtel 18-20, Wien

⇒ ImResFun: http://imresfun.cdl.univie.ac.at/finalmeeting/index.php

Publikation: Butyrate influences intracellular levels of adenine and adenine derivatives in the fungus Penicillium restrictum

04.2017

Zutz C, Chiang YM, Faehnrich B, Bacher M, Hellinger R, Kluger B, Wagner M, Strauss J, Rychli K (2017)

Microbiol Res. 2017 Apr;197:1-8. doi: 10.1016/j.micres.2016.12.013. Epub 2017 Jan 11.

Butyrate, a small fatty acid, has an important role in the colon of ruminants and mammalians including the inhibition of inflammation and the regulation of cell proliferation. There is also growing evidence that butyrate is influencing the histone structure in mammalian cells by inhibition of histone deacetylation. Butyrate shows furthermore an antimicrobial activity against fungi, yeast and bacteria, which is linked to its toxicity at a high concentration.

⇒ weiterlesen: http://dx.doi.org/10.1016/j.micres.2016.12.013

BiMM Poster Presentation at ÖGMBT Annual Meeting

12.-14.09.2016
8th ÖGMBT Annual Meeting – Life Sciences for the Next Generation – 2016, Graz, Austria.

Poster Title: The open research platform „BiMM -Bioactive microbial metabolites“ – a high-throughput biotic and chemical interaction approach to discover novel bioactive compounds

Abstract: Novel metabolic products are needed for health and biotechnology. Production of metabolites of axenic cultures has been exploited in the last decades. More recently, it has been realized that interactions of microorganisms can lead to production of dormant enzyme activities and even the exchange of metabolites between organisms can occur. Thus co-culture of microorganisms leads to a multitude of new biosynthetic possibilities. Moreover, mass sequencing results of microbial genomes predict a much richer diversity of microbial metabolites than previously anticipated. The exploitation of the potential of this microbial genetic dark matter has recently moved again into focus. Here we report on our approach to discover and characterize bioactive compounds and enzymes. We are using a variety of environmental and chemical conditions in which biotic interactions between different microbial genera trigger production of novel metabolites. Bioactivity testing of produced metabolites, enzymes, effectors and other compounds is performed in a high-throughput format using automated liquid and solid handling systems. Possible hits are followed up and characterized using a metabolomics platform developed at the Center for Analytical Chemistry. We have screened in a pilot study approximately 15k co-cultures of different bacteria and fungi combinations. Among these we identified about 30 interactions producing extracts with antibacterial and antifungal activity. Identification of the compounds responsible activity is under way.

The newly created HTP infrastructure is designed as open research platform providing hardware and expertise for dedicated collaborative projects in the field of innovative bioactive metabolite and enzyme production and characterization of their properties and function.

Authors:
Christoph Zutz 1, Christoph Schüller 2, Roland Hellinger 3, Kathrin Rychli 1, Martin Wagner 1, Rainer Schumacher 3, Scot Wallace 2, Joseph Strauss 2
Affiliation:
1 Institute of Milk Hygiene, Milk Technology and Food Science, VETMED
2 Department of Applied Genetics and Cell Biology, BOKU
3 Center for Analytical Chemistry, BOKU

Publikation: Valproic Acid Induces Antimicrobial Compound Production in Doratomyces microspores

13.04.2016

Zutz, C., Bacher, M., Parich, A., Kluger, B., Gacek-Matthews, A., Schuhmacher, R., … Strauss, J. (2016). Valproic Acid Induces Antimicrobial Compound Production in Doratomyces microspores. Frontiers in Microbiology, 7, 510.

One of the biggest challenges in public health is the rising number of antibiotic resistant pathogens and the lack of novel antibiotics. In recent years there is a rising focus on fungi as sources of antimicrobial compounds due to their ability to produce a large variety of bioactive compounds and the observation that virtually every fungus may still contain yet unknown so called “cryptic,” often silenced, compounds.

⇒ weiterlesen: http://journal.frontiersin.org/article/10.3389/fmicb.2016.00510/full

Bioactive Microbial Metabolites

Microorganisms and their metabolites have diverse impact on our lives. The goal of the research facility, Bioactive Microbial Metabolites (BiMM) is to isolate new active substances from fungi and bacteria.

The purpose of the BiMM is to provide an open research platform. The BiMM provides hardware and expertise for cooperation projects and contract work. The equipment is specially adapted to perform successful searches for new bioactive metabolites and enzymes. In addition, the BiMM is equipped to analyse the chemical properties and biological function of the found substances. We provide automated pipetting using modern liquid handling robots along with other automated operations. Another focus is the characterisation of chemical bioactive substances.

Slide 1

Slide 2