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Developing novel applications of mycelium based bio-composite materials for

design and architecture

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Building with bio-based materials: Best practice and performance specification

Developing novel applications of mycelium based bio-

composite materials for design and architecture
Noam Attias1,2, Ofer Danai2, Nirit Ezov2, Ezri Tarazi1, Yasha J. Grobman1
1
Industrial Design Department at the Faculty of Architecture and Town Planning, Technion -
Israel Institute of Technology, Haifa, 3200003, Israel, email: [email protected]
2
MIGAL - Galilee Research Institute, Israeli Northern R&D, Kiryat Shmona, 11016, Israel

Keywords: Bio-composite, Mycelium, Material, Design, Architecture

ABSTRACT

The growing need of the industry for alternative materials and products that are
biodegradable and derived from renewable resources has recently led researchers from
varied fields to search for more sustainable alternatives, and develop natural bio-
composites to replace varied petroleum-based products in order to reduce the intolerable
stress on the planet environment. In this research, the natural ability of saprophytic fungi
to bind and digest lingo-cellulose, is utilized to develop natural bio-composite materials
for novel applications in design and architecture.
Previous research has shown a potential to develop products such as packaging, building
and insulation materials, leather-like, textile and transparent edible films. However, no
research has been found in which all the significant variables were systematically tested.
In this work, several fungi species were grown on varied local agricultural-growth wastes
to evaluate which pair of fungi-plant material features the most suitable combination for
future applications. The fungi; Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus
salmoneostramineus and Aaegerita agrocibe where grown on woodchips of Eucalyptus,
Oak, Pine, Apple and vine. The samples were tested for selected properties, including
chemical changes in organic matter (pH, electric conductivity, water, carbon and nitrogen
contents), mycelium growth rate, density and quality impression. By examining these
fundamental materials characteristics, we aim to achieve a thorough understanding of the
structural and aesthetic opportunities that this novel bio-material should offer. The current
stage of the research shows that the most efficient integrations where the samples of P.
ostreatus grown on Apple or Vine woodchips. Future work will focus on using suitable
analytical methods for further understanding of the changes in mycelium and plant
structures during the digestion process, and locating essential variable parameters of
previous and post processing, to achieve desired material properties and introduce
innovative characteristics and functions over existing industrial products and applications.

INTRODUCTION

Utilizing fungi ability to digest varied lingo-cellulose wastes

Recent advances in technological, biological and digital computation abilities, alongside
the need for more sustainable manufacturing methods, assist to develop alternative
materials and fabrication processes, using multidisciplinary tools. In a world
characterized by rapid population growth, intensified agriculture and industrialization,
where fast, low-cost manufacturing process encourages a constant growth of production
and consumption, leading to an accumulation of waste, environmental pollution, and
depletion of natural resources, there is a growing need for alternative materials and
Building with bio-based materials: Best practice and performance specification

products, particularly in the mass industry, that are biodegradable and derived from
renewable resources.
A complex enzymatic process enables the white rot fungi with a unique ability to digest
highly stable molecules such as the structural polysaccharides of plants (Danai et al.
2012). This process plays a significant role in natural ecosystems and is already widely
used for varied agricultural and food applications (Levanon et al. 1988). However, the
fungal ability to bind and digest lignin and cellulose fibres of plants can also provide an
inherent bonding, forming a natural, light weight bio-composite, in which the fungal
mycelium functions as the matrix and holds the plant substrate pieces together, without
the use of any synthetic adhesives (Figure 7). The resulting material can therefore be
applied as a biodegradable alternative for a wide range of industrial materials, products
and applications.
In this paper, the novel concept of utilizing this integration of fungal mycelium with the
plant fibers, will be introduced. The paper starts with a general review of current design
works done with mycelium. It then presents a preliminary experiment we conducted, to
scan for a suitable fungi-plant set, as well as understand whether some analytical methods
can assist with the characterization of the final bio-composite material. It concludes that
there is a clear connection between the quantitative and qualitative parameters we have
tested so far. In future work, we wish to test more analytical methods to better characterize
the structural changes during mycelium development and its effect on the final material.
Using mycelium-plant material as a bio-composite in design and architecture
A bio-composite consisting of mycelium-plant material can be applied as a biodegradable
alternative for a wide range of industrial materials. At the end of its use, the mycelium
product can decompose to be available as a recourse for the development of other
organisms in the environment. Previous research has shown a potential to develop
products such as packaging (Holt et al. 2012 (Ziegler et al. 2016)), building (Jiang et al.
2016, 2013; Jiang et al. 2017; Travaglini et al. 2014), thermal and acoustic insulation
materials (Yang et al. 2014; M. G. G. Pelletier et al. 2013; M. G. Pelletier et al. 2017),
decorative house products (Mycoplast), leather (Mycoworks), textile (Hoitink) and
transparent edible films (Kumar 2000). In recent years, some workshops and exhibitions
(such as Fungal Futures 2016) of international artists and designers who investigate and
develop materials using mycelium are taking place. However, most of the current research
focus on evaluating the mechanical properties of the raw material on a macro level, using
a set of physical tests, such as compressive and tensile strength, stiffness, elasticity,
density, dimensional stability, aging, water absorption, thermal and acoustic insulation
(Mendez, Autónoma, and Antonio 2016; Lelivelt 2015). Yet, in order to detect and control
the final material properties, a bottom up approach is required, in which structural changes
within the plant and fungal components should first be identified. Using such approach
could contribute and advance the possible applications using mycelium based bio-
composites as novel materials.
Characterization of mycelium-plant bio-composite materials properties
The objective of this experiment is to locate the most suitable fungi-substrate combination
for further exploration and development of potential materials, products and applications
in the field of architecture and industrial design. In this work, four fungi species;
Pleurotus salmoneo-stramineus (P. salmoneo), Pleurotus ostreatus (P. ostreatus),
Pleurotus pulmonarius (P. pulmunarius), Aaegerita agrocibe (A. agrocibe); where grown
on five types of woodchips substrates; Eucalyptus ecamaldulensis (Eucalyptus); Quercus
calliprinos; (Oak), Pinus halepensis (Pine); Vitis vinifera (Vine- Cabernet Sauvignon);
Building with bio-based materials: Best practice and performance specification

Malus domestica (Apple- Golden Delicious). Each substrate was tested for selected
properties before and after the mycelium growth, including chemical changes in organic
matter (water, pH, electrical conductivity, ash, nitrogen content and organic matter
digestion); mycelium development was evaluated by rate, density and quality factors.
The water content was measured at the beginning of the experiment to test whether the
substrate contains a sufficient amount of water (about 65% is essential for fungal growth
and development); And at the end of the experiment, where it is expected to rise or remain
stable, due to the release of metabolic water during enzymatic digestion process.
The pH level of the substrate is expected to drop where fungal mycelium have developed.
A relatively high initial pH level (around 8) can donate to the selectivity of a substrate,
since Pleurotus mushrooms can manage to grow on higher pH levels than other, unwanted
fungi types. Furthermore, the digestion process of the fungi lowers the pH level of the
substrate to about 5), thus it might assist to indicate the amount of mycelium development
on the substrate.
The optimum nitrogen content range for Pleurotus mushroom growth is about 0.6-1%.
The organic matter composing wood contains about 50% carbon and around 1% nitrogen.
During its growth process, the mushroom digests mostly carbon from the substrate,
without consuming nitrogen. Thus, the relative amount of nitrogen in the final material
(substrate with mycelium) is expected to be higher compared with the control (only
substrate). The change in nitrogen content during mycelium development is therefore
another parameter to consider while evaluating mycelium development.
The amount of organic matter is determined by calculating the relative part of ash (non-
organic components) from the total mass of the initial substrate material, compared with
its amount on the final material (substrate with mycelium). Since the fungi digest organic
matter, the relative part of the ash will therefore increase, thus indicate the amount of
organic matter digested by the fungi. Hence, the loss of organic matter (∆OM) indicates
the amount of mycelium growth.
By accomplishing this experiment, we aim to understand which fungi-plant set is most
suitable to work with for design and architecture applications. Further experiments will
include a systematic testing for variations of substrate composition and characteristics,
while exploring advanced production methods to fully utilize the possibilities and
properties of this material. In future work, mycelium development could be evaluated
using quantitative visual inspections such as fluorescence chitin assay (Ayliffe et al.
2013). The unique fungal digestion mechanism could be utilized to manipulate and
enhance the mechanical properties of the plant fibres (Dresbøll and Magid 2006) to
feature integrated performances of the fungal mycelium that is bound within the plant
matter. As demonstrated in previous research (Avni et al. 2017), substrate composition
can also affect the structural components of fungal cell wall, therefore assist to manipulate
the final material properties.

EXPERIMENTAL

Mycelium cultivation
The mycelium cultivation process was done in the mushroom research lab of MIGAL,
northern R&D, Israel. The woodchips substrate for this experiment is pruning sourced
from local forest trees and agricultural crops in the Galilee area, Israel. The average
particle size is 5X15 mm. The substrate was thoroughly mixed with (50% w/v) water,
then placed in a ⌀14cm glass petri dish (150gr wet substrate in each plate) and sterilized
in an autoclave (1hr, 121°C). Each plate was inoculated with a disk of ⌀7mm mycelium
Building with bio-based materials: Best practice and performance specification

grown on agar, then incubated at 25°C for 4-5 weeks. Five replicas for each variation of
substrate type and fungi specie were used. Two plates of each substrate type (not
inoculated with mycelium) where used as control. Mycelium growth rate was calculated
by measuring its diameter every 2-3 days. Visual inspection was used to determine
mycelium quality and density, ranged from 1-Thin, almost transparent; to 5- thick and
firm, white (Figure 1). Mushroom growth parameters were multiply (growth rate mm/day
X mycelium density 1-5) and compared with organic matter loss to evaluate their relation.

B.
A.

Oak Pine Apple Vine Eucalyptus

A. agrocibe
P.pulmunarius

Figure 1: [A] Development of P. pulmonarius mycelium on oak for 16 days (from left to right). [B] Day
28, growth of A. agrocibe (top row) and P.pulmunarius (bottom row) on oak, pine, apple, vine and
eucalyptus substrates. mycelium density and growth rate were measured during the experiment.
Chemical parameters:
All chemical properties where tested before and after mycelium growth. Two repeats
where measured for each test. pH and conductivity where measured using a sample of
substrate soaked in distilled water (1:10 w/w). Water content was determined by oven-
drying (48 hours at 105°C). Samples where then milled using a 2mm sieve for carbon and
nitrogen tests. Nitrogen content determined according to Kjeldahl, using a Buchi K-435
digestion and B-324 distillation units. For ash content measurement, milled samples were
placed in crucible and burned in A Bifatherm multistage MS8 kiln (5 hours at 600°C).

Figure 2: Chemical parameters tests (from left to right); pH and electric conductivity, water, nitrogen
and ash contents.
Building with bio-based materials: Best practice and performance specification

RESULTS AND DISCUSSION

Water capacity
As shown in Figure 3, the initial water content of all selected substrates was 52-54%.
During the experiment, the control samples (without fungi) have lost significant amount
of water compared with mycelium containing samples. The P. pulmunarius samples
retained the highest water content during the experiment, followed by the P. ostreatus, P.
salmoneo and A. agrocibe (in this order). The growth of P. salmoneo and A. agrocibe on
vine and apple substrates did not develop or had a little growth and their water content
dropped similarly to the control. All types of fungi that were grown on the pine substrate
contained less water content compared with its control.

water capacity of substates

54
52
50
water [%]

48
46
44
42
40
Eucalyptus Vine Apple Pine Oak

control before control after A. agrocibe P. pulmunarius P. ostreatus P. salmoneo

Figure 3: The changes in water content of selected lignocellulosic substrates during growth of several
mushroom types. The mycelium retains higher water content compared with the control (before and
after the experiment).

The lack of current substrates to hold enough water seems to challenge the growth of
some fungi. Error! Reference source not found. demonstrates the effect of mycelium
development on retaining the water content in the substrate. The control was drier in
comparison to samples where mycelium has developed, except for the samples containing
pine substrate, which were all drier than its control. This might be related to the very low
density of mycelium on these samples (Figure 1). Furthermore, though the initial water
content of the apple substrate was the lowest, its combination with P. ostreatus and P.
pulmunarius retained its initial humidity and produced thick, dense mycelium.
Considering these results, it seems that changes in water content can point on the level of
mycelium development but does not necessarily indicate mycelium density.
pH level
As indicated in Figure 4, the P.ostreatus and P.salmoneo samples show a significant
decrease in pH level, yet all other results do not show a coherent correlation between pH
change and mycelium development. The initial pH level of all selected substrates was
around 5-5.5. Such pH level is not optimal for woodchips based substrates. This might
have influenced mycelium development.
Building with bio-based materials: Best practice and performance specification

Effect of fungi specie on pH level

7
6.5
6
5.5
pH

5
4.5
4
3.5
Eucalyptus Vine Apple Pine Oak

A. agrocibe P. pulmunarius P. ostreatus P. Salmoneo control before

Figure 4: Changes in pH level of selected lignocellulosic substrates during growth of several mushroom
types. Due to enzymatic digestion process, the pH level is expected to drop in comparison with the control
where fungal mycelium has developed.

Nitrogen content
Figure 5 indicates that in all sets, accept the A. agrocibe on oak, the nitrogen level
increased during mycelium development. The most significant increase is seen on the
eucalyptus substrate, that had a lower initial nitrogen content (0.6%) and increased during
the growth of P. ostreatus by 0.5%. On all other substrates, the change is smaller (about
2%). The oak substrate had a relatively high initial nitrogen content of (about 0.7%), yet,
the final values after the growth of varied fungi did not rise significantly (about 0.1%).

Effect of fungi specie on nitrogen content

1.2
1.1
1
0.9
%N

0.8
0.7
0.6
0.5
0.4
Eucalyptus Vine Apple Pine Oak

control before A. agrocibe P. pulmunarius P.ostreatus P. salmoneo

Figure 5: Changes in nitrogen content of selected lignocellulosic substrates during growth of several
mushroom types. Due to enzymatic digestion process, the relative nitrogen content is expected to increase
in comparison with the control.
Building with bio-based materials: Best practice and performance specification

Organic matter digestion

As can be seen in Figure 6, the samples of P.ostreatus grown on Vine and Apple substrates
and the P.pulmunarius on oak, have shown the highest loss of organic matter during
mycelium development (about 21gr). The smallest change is seen on the pine substrate.
The high correlation between the change in organic matter content and mycelium
development (Figure 7) indicate that this test is a reliable quantitative index to evaluate
mycelium quality.

Effect of fungi specie on organic matter digestion

25

20
∆OM [gr]

15

10

0
Eucalyptus Vine Apple Pine Oak

A. agrocibe P. pulmunarius P. ostreatus P. Salmoneo

Figure 6: Changes in organic matter content of selected lignocellulosic substrates during growth of
several mushroom types. Due to enzymatic digestion process, the relative organic matter content is
expected to decrease in comparison with the control. the largest change in organic matter is expected
where fungal growth is most developed.

Figure 7: P. ostreatus grown in ⌀ 14cm petri dishes with Apple (left) or Vine (right) woodchips.

Mushroom growth
To evaluate the mycelium quality and suitability for further work, mushroom growth
parameters were multiply (growth rate mm/day X mycelium density 1-5) and compared
with the amount of organic matter loss (Figure 8). This comparison shows high correlation
between the selected parameters (R squared value is close or same to 1). According to
this comparison, the mycelium growth on apple and vine substrates (Figure 7), showed
the most suitable qualitative and quantitative characteristics so far. According to this
Building with bio-based materials: Best practice and performance specification

comparison, there is a clear correlation between the quantitative change in organic matter
content during mushroom growth with the qualitative parameters of mycelium density
and thickness tested (Figure 1). Therefore, this comparison is the most reliable index to
evaluate mycelium development and suitability for further exploration so far.

Figure 8: A comparison between the amount of organic matter loss to mushroom growth parameters
(growth rate mm/day X mycelium density 1-5) shows high correlation, indicating it as reliable calculation
for mycelium development.

Substrate parameters
The results generally indicate a lack of few parameters that are essential for mushroom
growth and proper mycelium development. The water absorbance ability of desired
substrates should be increased to around 65%; the initial pH level of the substrate should
be higher, around 7-8; nitrogen content of the substrate should also start at higher levels,
closer to 1%. This could be achieved by a few actions:
a. Increase the surface area of the substrate components by reducing the particle size to
around Ø2mm as well as add varied amounts of sawdust (Danay et al. 2012), while
maintaining aerobic conditions in the substrate.
b. A prior heat treatment to open and expand air cavities between the wood fibers, thus
increase the porosity of the desired substrate.
c. Test additional materials with higher water absorbance ability, such as hemp fibers
(Lelivelt 2015), coconut, soy bean meal, wheat bran, gypsum etc. (Danay et al. 2012)
should be added to the selected substrate to increase its water capacity to the desired
amount. The addition of such substrates can also contribute to its initial nitrogen
content and to the mechanical properties (Lelivelt 2015).

CONCLUSIONS

The objective of this experiment is to locate fungi-substrate combinations that are most
suitable for further exploration and development of potential materials in the field of
architecture and industrial design. A set of 4 fungi species grown on 5 different
agricultural plant waste substrates. The samples were tested for selected properties,
Building with bio-based materials: Best practice and performance specification

including chemical changes in organic matter and qualitative evaluation of mycelium

development. The high correlation between organic matter loss and mushroom growth
parameters assisted to evaluate the efficiency of each substrate and fungi set. Best results
are P.ostreatus mycelium grown on vine and on apple substrates. Therefore, variations of
this fungi-plant sets could be used in further work. Yet, to better evaluate the suitability
of each substrate-fungi set for more particular applications, additional analytical methods
should be used. there is a need to develop a further set of parameters (physical, chemical
and visual) of the varied substrate components to understand their relations with one
another and its impact on final material properties.

ACKNOWLEDGEMENTS

This project was supported by the Faculty of Architecture and Town Planning at the
Technion, Israel institute of technology, in collaboration with the Edible Mushrooms
Department, Migal, northern R&D.

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