Page 15 - InnoRenew CoE International Conference 2023
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Time-lapse microscopic analysis of Aureobasidium spp.
growth and insights into environmental adaptation
Ana Gubenšek 1,3*, Aleksandar Tošić 1,4, Karen Butina Ogorelec 1,3,
Faksawat Poohphajai 1,2,3, Anna Sandak 1,3,4
1 InnoRenew CoE, Livade 6a, 6310 Izola, Slovenia, ana.gubensek@innorenew.eu, aleksandar.tosic@innorenew.eu, karen.butina@innorenew.eu,
faksawat.poohphajai@innorenew.eu, anna.sandak@innorenew.eu
2 Department of Bioproducts and Biosystems, School of Chemical Engineering, Aalto University, P.O. Box 16300, 00076 Aalto, Finland,
faksawat.poohphajai@aalto.fi
3 Andrej Marušič Institute, University of Primorska, Titov trg 4, 6000 Koper, Slovenia, ana.gubensek@iam.upr.si, karen.butina@iam.upr.si,
faksawat.poohphajai@iam.upr.si, anna.sandak@iam.upr.si
4 Faculty of Mathematics, Natural Sciences and Information Technologies, University of Primorska, Glagoljaška 8, 6000 Koper, Slovenia,
aleksandar.tosic@upr.si
* Corresponding author
The study of microorganisms, particularly fungi, at the microscopic level has gained increasing interest and
importance in recent years. Fungal populations and structures, such as biofilms, have become subjects of
investigation. Fungi initiate growth from spores, extending apical hyphae to form an interconnected, tree-
like mycelial network. This corded network exhibits species-specific variations and undergoes continuous
reconfiguration in response to environmental stimuli, involving new growth, branching, fusion, or regression.
Innovative methodologies, including time-lapse microscopic observation, have provided comprehensive
visualization and quantitative analysis of morphological features, characterisation of mycelial network
dynamics, and their response to environmental changes. The integration of time-lapse microscopic observation
with mathematical analysis offers valuable insights into simulating the growth of mycelia, which is crucial for our
understanding and controlling of fungal biofilm development, that can be implemented in various applications.
In this study, we focused on visualizing the growth of Aureobasidium spp. and evaluating their morphological
characteristics to generate statistical functions for simulating free mycelium growth. Cultures of A. pullulans
and A. melanogenum were observed using the EVOS™ M7000 Imaging System, which captured 3D and 2D
projection images at defined time intervals. The acquired images served as quantitative data for analysing
various morphological and growth parameters of Aureobasidium spp., such as tip growth rate, hyphal density,
branch angle, and branching location. Subsequently, we investigated the effects of nutrient, temperature, and
humidity variations on fungal growth. Moreover, the relationship between A. pullulans and A. melanogenum was
evaluated, as they are known to frequently colonize various building materials.
Keywords: Aureobasidium spp., growth, microscope, time-lapse observation
Acknowledgment: The authors gratefully acknowledge receiving funding from the European Union (ERC,
ARCHI-SKIN, #101044468). Views and opinions expressed are, however, those of the author(s) only and do
not necessarily reflect those of the European Union or the European Research Council. Neither the European
Union nor the granting authority can be held responsible for them. The authors gratefully acknowledge the
European Commission for funding the InnoRenew project (Grant Agreement #739574) under the Horizon2020
Widespread-Teaming program, the Republic of Slovenia (investment funding from the Republic of Slovenia and
the European Union’s European Regional Development Fund).
13–14 SEPTEMBER 2023 I IZOLA, SLOVENIA 15
growth and insights into environmental adaptation
Ana Gubenšek 1,3*, Aleksandar Tošić 1,4, Karen Butina Ogorelec 1,3,
Faksawat Poohphajai 1,2,3, Anna Sandak 1,3,4
1 InnoRenew CoE, Livade 6a, 6310 Izola, Slovenia, ana.gubensek@innorenew.eu, aleksandar.tosic@innorenew.eu, karen.butina@innorenew.eu,
faksawat.poohphajai@innorenew.eu, anna.sandak@innorenew.eu
2 Department of Bioproducts and Biosystems, School of Chemical Engineering, Aalto University, P.O. Box 16300, 00076 Aalto, Finland,
faksawat.poohphajai@aalto.fi
3 Andrej Marušič Institute, University of Primorska, Titov trg 4, 6000 Koper, Slovenia, ana.gubensek@iam.upr.si, karen.butina@iam.upr.si,
faksawat.poohphajai@iam.upr.si, anna.sandak@iam.upr.si
4 Faculty of Mathematics, Natural Sciences and Information Technologies, University of Primorska, Glagoljaška 8, 6000 Koper, Slovenia,
aleksandar.tosic@upr.si
* Corresponding author
The study of microorganisms, particularly fungi, at the microscopic level has gained increasing interest and
importance in recent years. Fungal populations and structures, such as biofilms, have become subjects of
investigation. Fungi initiate growth from spores, extending apical hyphae to form an interconnected, tree-
like mycelial network. This corded network exhibits species-specific variations and undergoes continuous
reconfiguration in response to environmental stimuli, involving new growth, branching, fusion, or regression.
Innovative methodologies, including time-lapse microscopic observation, have provided comprehensive
visualization and quantitative analysis of morphological features, characterisation of mycelial network
dynamics, and their response to environmental changes. The integration of time-lapse microscopic observation
with mathematical analysis offers valuable insights into simulating the growth of mycelia, which is crucial for our
understanding and controlling of fungal biofilm development, that can be implemented in various applications.
In this study, we focused on visualizing the growth of Aureobasidium spp. and evaluating their morphological
characteristics to generate statistical functions for simulating free mycelium growth. Cultures of A. pullulans
and A. melanogenum were observed using the EVOS™ M7000 Imaging System, which captured 3D and 2D
projection images at defined time intervals. The acquired images served as quantitative data for analysing
various morphological and growth parameters of Aureobasidium spp., such as tip growth rate, hyphal density,
branch angle, and branching location. Subsequently, we investigated the effects of nutrient, temperature, and
humidity variations on fungal growth. Moreover, the relationship between A. pullulans and A. melanogenum was
evaluated, as they are known to frequently colonize various building materials.
Keywords: Aureobasidium spp., growth, microscope, time-lapse observation
Acknowledgment: The authors gratefully acknowledge receiving funding from the European Union (ERC,
ARCHI-SKIN, #101044468). Views and opinions expressed are, however, those of the author(s) only and do
not necessarily reflect those of the European Union or the European Research Council. Neither the European
Union nor the granting authority can be held responsible for them. The authors gratefully acknowledge the
European Commission for funding the InnoRenew project (Grant Agreement #739574) under the Horizon2020
Widespread-Teaming program, the Republic of Slovenia (investment funding from the Republic of Slovenia and
the European Union’s European Regional Development Fund).
13–14 SEPTEMBER 2023 I IZOLA, SLOVENIA 15
