Copyright © 2012 American Scientific Publishers Journal of

All rights reserved Biobased Materials and Bioenergy

Printed in the United States of America Vol. 6, 431–439, 2012

Fungal Mycelium and Cotton Plant Materials in the

Manufacture of Biodegradable Molded Packaging
Material: Evaluation Study of Select
Blends of Cotton Byproducts
G. A. Holt1 ∗ , G. McIntyre2 , D. Flagg2 , E. Bayer2 , J. D. Wanjura1 , and M. G. Pelletier1

RESEARCH ARTICLE
1
USDA-ARS Cotton Production and Processing Research Unit 1604 E. FM 1294, Lubbock, TX 79403, USA
2
Ecovative Design, LLC, 60 Cohoes Avenue, Green Island, NY 12183, USA

Delivered
Since polystyrene is non-biodegradable, by Ingenta
a biodegradable to: that is eco-friendly is being sought
material
as a substitute for packaging and insulation Greg
boardHolt
consumers. One such process, developed by
Ecovative Design, LLC, involves growingIP fungal
: 199.133.133.66
species on agricultural biomass to produce an eco-
friendly packaging product (EcoCradle™)
Thu, 29 andNovinsulation panels (Greensulate™). The objective of
2012 23:20:57
this research was to develop and evaluate six blends of processed cotton plant biomass (CPB)
materials as a substrate for colonization of selected fungi in the manufacture of molded packaging
material. The blends were comprised of processed CPB, cotton seed hulls, starch, and gypsum. The
four ingredients were the same mix percentage for all six blends with the particle size of the CPM
being the only difference. CPB particles sizes ranged from 0.1 to 51 mm. Tests were conducted to
evaluate the physical and mechanical properties of the six CPB blends. Test results revealed blends
that met or exceeded like characteristics of extruded polystyrene foam.
Keywords: Composite, Cotton, Mycelium, Biobased, Biodegradable.

1. INTRODUCTION studied and used to manufacture environmentally friendly

Polystyrene is a hydrocarbon based material typically
used in the manufacture of packaging materials. Marketed
under the name Styrofoam™, this lightweight material
is hydrophobic, resistant to photolysis, and is not sub-
ject to decomposition or decay.1 These characteristics are
attractive to shippers and the packaging industry, but they
create problems with respect to recycling, reuse, and land-
fill operation.2 Recent scientific investigation resulted in
development of an economically viable, environmentally
friendly replacement for polystyrene packaging materials.
The packaging material evaluated in our study is a com-
posite containing selected agricultural residues and a spe-
cific fungus.
Annually renewable crops and their agricultural residues
have been researched extensively as materials that could
potentially enhance the properties of composite products
made from fossil fuel based derivatives that are more resis-
tant to biodegradation.3–6 In addition to the crops and agri-
cultural residues, fungi and/or their constituents have been
∗
Author to whom correspondence should be addressed.
Email: greg.holt@ars.usda.gov
products.7–10 The combination of agricultural residues and
fungi have been evaluated for fungal cultivation11–15 and
improving bonding properties of agricultural fibers in the
manufacture of composites.16 17 However, a method for
producing a composite comprised of agricultural residues
and fungi was not found in the literature.
Bayer et al.18 and Bayer and McIntyre19 developed pro-
cesses that involve growing fungal species on agricul-
tural residues, such as cotton plant material, to produce an
environmentally-friendly packaging material. The objec-
tive of the present study was to develop six blends of
specifically processed cotton plant material for use with
the Bayer et al.18 and Bayer and McIntyre19 processes.
Each of the six blends was used to produce a packag-
ing material that was subjected to standard test methods
for compressive strength,20 flexural strength,21 modulus of
elasticity,21 density,22 dimensional stability,22 accelerated
aging,23 water absorption,24 cone calorimetry,25 and ther-
mal conductivity.26 The cotton plant material used in this
study was a byproduct of typical mechanical harvesting
and ginning practices in the United States which gener-
ate approximately 2.5 Mg of cotton byproducts across the
U.S. cotton belt each year.27

J. Biobased Mater. Bioenergy 2012, Vol. 6, No. 4 1556-6560/2012/6/431/009 doi:10.1166/jbmb.2012.1241 431

 Fungal Mycelium and Cotton Plant Materials in the Manufacture of Biodegradable Molded Packaging Material
2. MATERIALS AND METHODS
2.1. Cotton Plant Biomass and Blends
The cotton plant material was obtained from a cotton gins
agricultural waste stream. Specifically, the cotton carpel
(bur) was obtained from the extractors27 in the United
States Department of Agriculture–Agricultural Research
Service (USDA-ARS) gin lab in Lubbock, Texas. The
extractors removed approximately 55% of the total waste
stream, primarily sticks and carpel, from the incoming
seedcotton before the seedcotton entered the gin stand(s).
The cotton carpels were processed through a Jacobson
model 4D hammer-mill (Jacobson Machine Works, Min-
neapolis, MN) or a Reynolds Engineering model T00-18
attrition mill (Reynolds Engineering and Equipment,
RESEARCH ARTICLE
Muscatine, IA) and then sorted across a Triple/S Dynam-
ics model VCSUF-24X12-3 vibratory conveyor (Triple/S
Dynamics, Inc., Dallas, TX) to obtain the specific particle
size ranges needed for each blend (Table I).Delivered
carpel material, for one of the blends, before and after
processing is shown in Figure 1. The sized material
packaged and stored in a dry location until Thu, blending.29 Nov
cotton plant material particles were sized to be within the
desired range, 0.1 to 51 mm.
Blending of the cotton plant material was accomplished
with a Davis 1.4 Mg ribbon mixer (H.C. Davis Sons Man-
ufacturing Co., Inc., Bonner Springs, KS). Each blend was
processed in 159 kg batches. The constituent materials
used in the six blends evaluated were comprised of:
(1) processed cotton carpel,
(2) cotton seed hull,
(3) starch, and
(4) gypsum.
The primary ingredient in each of the six blends was the
processed cotton carpel. For all six blends, the ingredi-
ents were added at the same percentages with the only
difference being the particle size range of the processed
cotton carpel (Table I). Each ingredient was added one at
a time into the mixer and agitated while adding between
1.5 to 2.5 l of water, to minimize dust and promote
adhesion of the starches and gypsum to the cotton plant
Table I. Particle size ranges for cotton plant material used in each of
the six blends evaluated in this study.
Primary particle size range of
Blends a
processed cotton plant material (mm)
1 28–51
2 12–28
3 0.1–12
4 12–51
5 0.1–12, 28–51
6 0.1–51
Notes: a Primary ingredient of each blend was the processed cotton plant material. listed are considered proprietary information by Ecovative
The other ingredients in each blend were starch, gypsum, and cottonseed hulls.
432
Holt et al.
Fig. 1. Some of the cotton plant material used in this study before (left)
and after (right) processing.
material. Once all the ingredients were added, the blend
was allowed to mix for 7 min, at maximum agitation
(15 rpm). Upon completion of the mixing, the blend
was emptied into a tote bag, labeled, and stored until
The cotton by Ingenta
shipmentto: to Ecovative Design’s facility in Green Island,
Greg Holt
NY. Specifics related to constituent fraction of the blends
IP : 199.133.133.66
was are considered proprietary information by Ecovative
The 2012 23:20:57
Design, LLC.
2.2. Composite Fabrication
At Ecovative’s research laboratory in Green Island,
New York, is the pilot manufacturing plant. A schematic
of the pilot plant process is shown in Figure 2. At the
pilot plant, the tote bags with each blend were emptied
into a bulk bin with a live-bottom auger that fed a pas-
teurizer where the material was sterilized at 115  C for
approximately 28 min. Exiting the pasteurizer, the blend
was gravity fed into a water-jacketed auger (cooler) where
it was cooled below 35  C. Upon exiting the cooler, the
blend was inoculated with the fungus, Ganoderma sp.,
using a specified grain or liquid substrate as the carrier.
After inoculation, the blend was discharged into a plas-
tic mold, referred to as a tool, which was in the desired
shape of the piece to be fabricated. The material was gen-
tly hand-packed in the tool and any excess was removed,
the tool was then sealed in order to maintain a consistent
micro-environment for fungal propagation (Fig. 3). The
filled tool was incubated on a bread rack at 21  C for
5-days at which time the fungal mycelium colonized the
blend. Figure 4 shows the fungal colonization of one of
the blends over a three day period. After 5 days, the part
was removed from the tool and placed in 60  C convec-
tion oven for 8 h, which inactivated the fungus and pre-
vented reanimation. After drying, the pieces were stored at
ambient laboratory conditions (approximately 21  C and
30% RH) until testing. Figure 5 shows a typical part after
drying. Specifics related to quantity of inoculum, pasteur-
izer and cooler speeds, and specific processes applied not
Design, LLC.
J. Biobased Mater. Bioenergy 6, 431–439, 2012
Holt et al. Fungal Mycelium and Cotton Plant Materials in the Manufacture of Biodegradable Molded Packaging Material

RESEARCH ARTICLE
Delivered by Ingenta to:
Greg Holt
IP : 199.133.133.66
Thu, 29 Nov 2012 23:20:57

Fig. 2. Schematic of Ecovative’s pilot manufacturing facility used to produce the cotton plant and fungal mycelium based molded packaging specimens
evaluated in this study.

2.3. Analytical Testing
Each of the six blends was used to produce a packag-
ing material that was subjected to standard test meth-
ods for compressive strength,20 flexural strength21 (Fig. 6),
modulus of elasticity,21 density,22 dimensional stability,22
accelerated aging,23 water absorption,24 cone calorimetry25
(Fig. 7), and thermal conductivity.26 Cone calorime-
try (flame retardance characteristics) was performed at
the Worcester Polytechnic Institute (WPI) Fire Research
Laboratory in Worcester, MA. Specimens were tested in
horizontal orientation at 50 kW/m2 heat flux. All other
analyses were conducted at the Ecovative research labo-
ratory. We did not evaluate expanded polystyrene samples
in this study. Numerous sources of information pertain-
ing to physical and mechanical properties of expanded
polystyrene are available in the public domain.28–31
2.4. Data Analysis
Two types of inoculum (grain and liquid substrate) were
applied to each of six cotton plant material blends for a

Fig. 3. Selecting the lid for the tool containing inoculated cotton plant material substrate (left) and snapping the lid in place (right) to maintain
micro-environment for optimum growth.

J. Biobased Mater. Bioenergy 6, 431–439, 2012 433

 Fungal Mycelium and Cotton Plant Materials in the Manufacture of Biodegradable Molded Packaging Material Holt et al.

Fig. 5. Finished fungal mycelium molded packaging piece after drying.

total of 12 treatments. Each treatment was replicated from

RESEARCH ARTICLE

3 to 12 times depending on the test method and property

being evaluated. Standard analysis of variance techniques
were used to analyze the data to determine statistically
Delivered by Ingenta
significantto:differences among the 12 treatments by the
Greg Holt
Tukey-Kramer Honestly Significant Difference (HSD) test
IP : 199.133.133.66
(release 9.2, SAS Institute Inc., Cary, NC) at the 95% con-
Thu, 29 Nov 2012 23:20:57
fidence level.

Fig. 6. Flexural strength testing of one of the cotton plant material test
specimens.

Fig. 4. Time sequence showing the inoculated cotton plant material sub-
strate (day 0) and fungal colonization over a 3-day period. The white Fig. 7. Cone calorimeter test samples of two of the treatments evaluated
specks are the living fungus. in this study, Grain 6 (left) and Grain 4 (right), after testing.

434 J. Biobased Mater. Bioenergy 6, 431–439, 2012

Holt et al. Fungal Mycelium and Cotton Plant Materials in the Manufacture of Biodegradable Molded Packaging Material

3. RESULTS AND DISCUSSION
3.1. Physical Properties
Figure 8 shows a comparison of the dimensional stability
(surface area contraction) of the treatments after drying.
Blends inoculated with the grain-based substrate showed
less surface area contraction than did blends inoculated
with the liquid-based substrate. Grain 3 (Blend 3, grain-
based inoculum) had the smallest measured surface area
contraction (0.64%), whereas Liquid 5 (Blend 5, liquid-
based inoculum) exhibited the greatest measured surface
area contraction (2.4%). Grain 3, 4, 5, and Liquid 6 were
similar in the percent contraction and significantly lower
than Grain 2, Liquid 1, 2, 3, 4, and 5.
The importance of dimensional stability is related to
Degradation Percentage
due to the greater mass of the grain-based inoculum versus
the liquid-based inoculum. The density adjusted values for
FS show Grain 1 and Grain 6 with the highest values at
26 kPa and Liquid 2 with the lowest at 7 kPa. The EM
was significantly higher for Liquid 5 (674 kPa) than all
other treatments with Liquid 2 having the lowest, 123 kPa.
Compressive strength was significantly higher for Liquid 3
(72 kPa) than for all other treatments with Liquid 4 having
the lowest CS at 1.1 kPa.
Sample degradation associated with FS, EM, and CS
resulting from accelerated aging testing is shown in
the second column of Table II. The percent degrada-
tion data was calculated according to the equation in the
standard:23

RESEARCH ARTICLE
tool design. The larger the percent contraction the more
oversized the tool needs to be for the finished product to = (Conditioned test value/as received test value) ∗ 100
be within desired specifications. Another factor influenc-
ing tool design is contraction variability. TheDelivered
more vari- by Ingenta to: values closer to the base line (100%) exhibit
Therefore,
able a blend/inoculum combination is, the more difficult Greg Holt
less degradation than samples with values further from the
it is to produce parts that are consistently withinIPdimen-
: 199.133.133.66
baseline. FS degradation for Grain 5, 1, 6, and Liquid 3
Thu,
sional tolerances of customer specifications. All 29 Nov 2012
treatments 23:20:57
exhibited little degradation from aging. Liquid 5’s FS was
had similar standard mean errors associate with percent reduced almost in half as a result of aging whereas the
contraction (0.093 to 0.108), so the means are a reliable FS of Grain 3 and Liquid 2 exhibited increased stiffness
indicator of the contraction expected when designing tools due to aging. EM for Grain 6 had the largest change in
for a given treatment. percent degradation of 318.6%. Liquid 5 had the largest
The flexure strength (FS), elastic modulus (EM), and reduction in EM at 43.6%. The treatments that had the
compressive strength (CS) in Table II are normalized to largest percent CS degradation were Liquid 5 (250.1%)
a standard density of 32.04 kg/m3 since this is the den- and Liquid 3 (6.8%). The CS degradation was least for
sity of the polystyrene packaging the EcoCradle material Grain 4 (92%) and Grain 1 (110%). Overall, Grain 1 had
can replace in the market. The density of the treatments the most consistent performance, by exhibiting some of the
ranged from 66.5 kg/m3 to 224 kg/m3 . The density for lowest degradation values for FS, EM, and CS compared
grain treatments was higher than for the liquid treatments to all other treatments.

Fig. 8. Average surface area contraction (%) or shrinkage of the sample pieces made from each treatment after oven drying. Bars with the same
letters are not significantly different at the 0.05 level of significance.

J. Biobased Mater. Bioenergy 6, 431–439, 2012 435

 Fungal Mycelium and Cotton Plant Materials in the Manufacture of Biodegradable Molded Packaging Material Holt et al.

Table II. Physical properties from molded packaging material produced Table III. Water absorption testing results from molded packaging
from six cotton-based substrate blends and two fungal inoculum methods. material produced from six cotton-based substrate blends and two fungal
inoculum methods.
Physical properties (density adjusted)
Water absorption
Response variablea Flexure strength Flexure strength degradation
Standardb ASTM C203 ASTM C481 Response Gain after Gain after Gain after
Units (kPa) (%) variable 75 hr 3 hr 168 hr
Standard ASTM C1134 ASTM C1134 ASTM C1134
Inoculum/blend Valuec Inoculum/blend Valuec
Units (%) (%) (%)
Grain 1 261A Liquid 2 1967A
Inoculum/ Inoculum/ Inoculum/
Grain 6 259A Grain 3 1860A blend Valueb blend Valueb blend Valueb
Grain 2 235A B Grain 4 1812A B
Grain 5 219A B C Liquid 4 Grain 1
1583A B 307A Grain 1 480A Grain 1 1981A
Liquid 1 208A B C Liquid 6 Liquid 1
1557A B 207A B Liquid 4 392A B Liquid 4 1780A
Liquid 5 196A B C Liquid 1 Grain 5
1151A B 171B C Liquid 1 342A B Liquid 1 1688A
Liquid 2 139B C Grain 5 298A B Liquid 5 1637A
Liquid 3 164A B C Grain 2 1116A B
Liquid 5 115B C Liquid 5 258A B Grain 5 1550A
Grain 3 123B C Grain 6 1005A B
Liquid 4 112B C Liquid 2 252A B Liquid 2 1536A
Grain 4 104B C Grain 1 1003A B
RESEARCH ARTICLE

Liquid 3 103B C Liquid 6 214A B Liquid 6 1324A

Liquid 6 101B C Grain 5 1001A B
Grain 2 99B C Grain 3 179B Grain 2 1269A
Liquid 4 97B C Liquid 3 950A B
Grain 3 93B C Grain 2 177B Grain 4 1103A
Liquid 2 70C Liquid 5 600BGrain 4 90B C Liquid 3 155B Liquid 3 1061A
Response variablea Elastic modulus
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Elastic modulus degradation Liquid 6 to: 70C Grain 4 122B Grain 6 943A
Standardb ASTM C203 ASTM C481 Greg Holt
Grain 6 64C Grain 6 108B Grain 3 935A
Units (kPa) (%) IP : 199.133.133.66
Notes: ASTM = American Society for Testing and Materials. b Means
a
within the
c
Thu, 29 Nov
c
2012
same 23:20:57
column followed by different letters in the corresponding row are statistically
Inoculum/blend Value Inoculum/blend Value different at the 0.05 level of significance.
Liquid 5 6745A Grain 6 3186A
Grain 2 3956B Liquid 2 1894A B
Grain 1 3681B C Grain 3 1659A B
3.2. Water Absorption
Liquid 3 3442B C Grain 5 1641B
The water absorption results (Table III) show the percent
Liquid 4 3318B C Grain 4 1632A B
Grain 3 3174B C Liquid 6 1507A B weight gain after 0.75 h, 3 h, and 168 h. After 0.75 h, Grain
Liquid 6 3088B C Liquid 3 1421B 6 (6.4%) and Liquid 6 (7.0%) had the lowest water absorp-
Grain 6 2884B C Grain 2 1347A B tion whereas Grain 1 (30.7%) and Liquid 1 (20.7%) had the
Grain 5 2834B C Liquid 4 1219B highest. Grain 1 had significantly higher water absorption
Liquid 1 2538B C Grain 1 984B after 0.75 h than all other treatments except Liquid 1.
Grain 4 1991B C Liquid 1 955B
Liquid 2 1228C Liquid 5 436B
Table IV. Conductivity and R-values from molded packaging material
Response Compressive Compressive strength produced from six cotton-based substrate blends and two fungal inoculum
variablea strength degradation methods.
Standardb ASTM C165 ASTM C481
Units (kPa) (%) Thermal properties

Inoculum/blend Valuec Inoculum/blend Valuec Response variable R-value Conductivity

Standarda TPS500 TPS500
Liquid 3 722A Liquid 5 2501A Units – W/m ∗ K
Liquid 1 335B Grain 3 1695A B
Inoculum/blend Valueb Inoculum/blend Valueb
Grain 6 110B C Grain 2 1399A B
Grain 2 90B C Liquid 4 1202A B Liquid 6 151A Grain 3 018A
Grain 5 85B C Grain 1 1101A B Liquid 3 140A B Liquid 4 017A
Grain 4 62C Grain 4 917A B Liquid 1 128A B Grain 1 016A B
Grain 1 54C Liquid 6 877A B Grain 2 115A B Liquid 5 015A B
Liquid 2 33C Grain 6 605A B Grain 4 114A B Grain 6 015A B
Liquid 6 30C Liquid 2 477A B Grain 5 112A B Liquid 2 015A B
Grain 3 23C Grain 5 421A B Liquid 2 105A B Grain 5 013A B
Liquid 5 14C Liquid 1 81A B Grain 6 100A B Grain 2 013A B
Liquid 4 11C Liquid 3 68B Grain 1 096B Grain 4 013A B
Liquid 4 087B Liquid 1 012A B
Notes: a Response variables are normalized to a density of 32.04 kg/m3 . Liquid 5 084B Liquid 3 012A B
Degradation = (Tested specimen value/as received specimen value) ∗ 100. Degrada- Grain 3 082B Liquid 6 010B
tion values of 100 indicate no change before and after testing. b ASTM = American
Society for Testing and Materials. c Means within the same column followed by Notes: a TSP = Transient plane source from therm test incorporated, Frederiction,
different letters in the corresponding row are statistically different at the 0.05 level Nebraska, USA. b Means within the same column followed by different letters in
of significance. the corresponding row are statistically different at the 0.05 level of significance.

436 J. Biobased Mater. Bioenergy 6, 431–439, 2012

Holt et al. Fungal Mycelium and Cotton Plant Materials in the Manufacture of Biodegradable Molded Packaging Material

After 3 h, Grain 1 (48.0%) still had the highest water The largest increase in water absorption from 0.75 h to 3 h
absorption and Grain 6 (10.8%) the lowest. However, was seen in Liquid 4 which moved from 11.2% to 39.2%.
Grain 1 was only significantly higher than five other treat- The smallest increase from 0.75 h to 3 h was Grain 4,
ments: Grain 2, Grain 3, Grain 4, Grain 6, and Liquid 3. 8.9% to 12.2%.

Table V. Cone calorimeter testing results from molded packaging material produced from six cotton-based substrate blends and two fungal inoculum
methods.

Thermal properties

Response variable Peak heat release rate Average heat release rate Peak carbon monoxide
Standarda ASTM E1354 ASTM E1354 ASTM E1354
Units kW/m2 kW/m2 kW/m2
Inoculum/blend Valueb Inoculum/blend Valueb Inoculum/blend Valueb

Liquid 2 1158A Liquid 2 753A Grain 6 5550A

Grain 1 1133A Liquid 6 665A B Liquid 2 247B

RESEARCH ARTICLE
Liquid 6 1070A Grain 5 663A B Liquid 4 229B
Grain 4 1053A Liquid 4 625A B Liquid 6 213B
Grain 2 1040A Grain 3 617A B Grain 1 200B C
Grain 3 1040A Liquid 5 613A B Grain 5 200B C
Grain 6 1035A DeliveredGrain
by4Ingenta to:593A B Liquid 5 197B C
Grain 5 1033A Grain Holt
Greg 2 572A B Liquid 1 177B C D
Liquid 5 991A Grain 6 565A B Grain 4 167B C D
Liquid 4 973A B
IP : 199.133.133.66
Liquid 1 562A B Grain 2 158B C D
Liquid 1 919A B Thu, 29 Nov
Grain2012
1 23:20:57
553B Grain 3 100C D
Liquid 3 657B Liquid 3 549B Liquid 3 090D

Response variable Average carbon monoxide Peak carbon dioxide Average carbon dioxide
Standarda ASTM E1354 ASTM E1354 ASTM E1354
Units mg/s mg/s mg/s
Inoculum/blend Valueb Inoculum/blend Valueb Inoculum/blend Valueb

Grain 6 5100A Grain 6 26180A Grain 6 2 4280A

Liquid 6 077B Liquid 6 4014B Liquid 2 2465B
Grain 2 068B C Grain 4 3963B Liquid 6 2203B
Liquid 4 052B C Grain 1 3897B Liquid 4 2098B
Liquid 2 050B C Grain 5 3827B C Grain 5 2027B
Liquid 5 046B C Liquid 2 3826B C Liquid 5 1976B
Liquid 1 044B C Liquid 5 3412B C Grain 4 1957B
Liquid 3 034B C Liquid 1 3235B C Grain 1 1863B
Grain 3 033B C Grain 3 3127B C Liquid 1 1856B
Grain 1 000C Liquid 4 3119B C Grain 3 1723B
Grain 4 000C Grain 2 3101B C Liquid 3 1516B
Grain 5 000C Liquid 3 2378C Grain 2 1446B

Response variable Peak mass loss rate Average mass loss rate Mass loss
Standarda ASTM E1354 ASTM E1354 ASTM E1354
Units mg/s mg/s mg/s

Inoculum/blend Valueb Inoculum/blend Valueb Inoculum/blend Valueb

Grain 1 1533A Liquid 3 714A Liquid 4 740A

Grain 4 1500A B Liquid 6 667A Grain 5 727A
Liquid 4 1367A B Liquid 5 621A B Grain 1 720A
Grain 5 1267A B Liquid 4 616A B Grain 2 702A
Grain 6 1250A B Liquid 1 591A B Grain 6 700A
Liquid 6 1200A B Grain 6 520A B Grain 4 690A
Grain 2 1150A B Grain 2 460A B Liquid 2 686A
Liquid 1 1133A B Grain 1 387A B Liquid 5 555A
Liquid 5 1067A B Grain 5 370A B Grain 3 527A B
Grain 3 1033A B Grain 3 363A B Liquid 1 462A B
Liquid 3 900A B Grain 4 170A B Liquid 6 396A B
Liquid 2 767B Liquid 2 15B Liquid 3 88B

Notes: a ASTM = American Society for Testing and Materials. b Means within the same column followed by different letters in the corresponding row are statistically
different at the 0.05 level of significance.

J. Biobased Mater. Bioenergy 6, 431–439, 2012 437

 Fungal Mycelium and Cotton Plant Materials in the Manufacture of Biodegradable Molded Packaging Material Holt et al.

The percent water absorption after 168 h indicated no

significant differences between any of the treatments. The
largest absorption, after 168 h, was seen in Grain 1 (198%)
with the lowest percent absorption in Grain 3 and Grain 6,
93.5% and 94.3%, respectively.

3.3. Thermal Properties

The thermal properties (Tables IV and V) show thermal
conductivity26 and cone calorimetry25 data. The thermal
resistance and conductivity data (Table IV) show Liq-
uid 6 with the highest R-value (1.5) which was signifi-
cantly higher than Grain 1 (0.96), Liquid 4 (0.87), Liquid 5 Fig. 9. Fungal mycelium and cotton plant material molded packaging
(0.84), and Grain 3 (0.82). Thermal conductivity was high- material being used by a large office equipment manufacturer in the
United States.
est with Grain 3 (0.18) and lowest with Liquid 6 (0.10).
The conductivity values were within the ranges of gypsum
RESEARCH ARTICLE

(0.17), high density hardboard (0.15), plywood (0.12), and

both hardwoods (0.16) and softwoods (0.12) at 26.8  C.32
Cone calorimeter testing (Table V) showed gas pro-
Delivered by Ingenta to:
duction (carbon monoxide, CO and carbon dioxide, CO2 )
for Grain 6 to have significantly higher gas production
Greg Holt
IP
than all other treatments, 55.5 mg/s—peak CO; 51 mg/s—
: 199.133.133.66
Thu, 29 Nov 2012 23:20:57
average CO; 2618 mg/s peak CO2 ; 2428 mg/s—average
CO2 . Originally upon examination of the large difference
between the Grain 6 data and the other treatments, the
Grain data was questioned as a potential error in data entry
or perhaps a mistake during the analytical analyses. Upon
further review and analysis, validity of the data was estab-
lished by investigation into the interaction of substrate par-
Fig. 10. Fungal mycelium and cotton plant material molded packag-
ticle size and method of inoculation to cone calorimeter ing material being used by a large computer manufacturer in the United
analysis. The lowest peak CO gas production was seen States.
in Liquid 3 (0.9 mg/s). The average CO production for
Grain 1, Grain 4, and Grain 5 show zero emissions when in
an application using treatments Grain 1 and Grain 2 and
actuality they are less than 0.00 mg/s. With the exception
Figure 10 is a hybrid whose development was a direct
of Grain 6, all other treatments had average CO production
result of the findings in this study.
less than 1.0 mg/s. The peak and average CO2 production
was lowest for Liquid 3 (237.8 mg/s—peak; 151.6 mg/s—
average) and Grain 2 (310.1 mg/s—peak; 144.6 mg/s— 4. CONCLUSIONS
average).
Cone calorimeter testing showed Liquid 3 had the small- This study evaluated six cotton-based biomass blends for
est mass loss (8.8%) compared to Liquid 4 with 74%. use in a process designed to produce an environmentally-
Liquid 3 had significantly lower mass loss than all treat- friendly molded packaging material that could replace
ments except Grain 3 (52.7%), Liquid 1 (46.2%), and polystyrene packaging currently in the marketplace.
Liquid 6 (39.5%). The peak and average mass loss rate In addition to the six cotton-based blends, two methods,
were lowest for Liquid 2, 76.7 mg/s and 1.5 mg/s, respec- grain and liquid, were used to inoculate the blends with
tively. The highest peak and average mass loss rates fungal spores resulting in twelve treatments. The blends
were seen in treatments Grain 1 (153.3 mg/s—peak) and were inoculated and the test specimens grown in tools
Liquid 3 (71.4 mg/s—average). (plastic molds) for 5 days and then dried to remove mois-
ture. The recipe for each blend was identical except for
3.4. Application of Findings the particle size range of the cotton-based materials. The
difference in the inoculums was that one carried the fungal
Results of this study indicate that cotton-based fungal spores on kernels of grain whereas the other had the fungal
mycelium packaging material is a viable alternative to spores suspended in liquid. The liquid inoculum was easier
polystyrene packaging. Figures 9 and 10 show two com- to use in the process and provided a more consistent dis-
mercial applications for the type of products produced tribution of fungal spores when applied to the blends. The
from two blends evaluated in this study. Figure 9 shows grain inoculum generally resulted in higher densities due

438 J. Biobased Mater. Bioenergy 6, 431–439, 2012

Holt et al. Fungal Mycelium and Cotton Plant Materials in the Manufacture of Biodegradable Molded Packaging Material
primarily to the added weight of the grain. The densities
were higher than desired (32.04 kg/m3 ) due in large part
to the inclusion of cotton plant particles less than 2 mm.
In future studies, cotton plant material having a diame-
ter less than 2 mm will not be used. No single treatment
outperformed the other treatments in all categories evalu-
ated. Most of the treatments performed similarly to each
other for the response variables measured. In regards to
percent degradation associated with accelerated aging test-
ing, Grain 1 was most consistent in maintaining flexural
and compressive strength and elastic modulus.
Overall, the use of cotton-based fungal mycelium pack-
aging material is a viable alternative to polystyrene
packaging. As refinements in processing and biomass
blend development continue, the physical and mechanical
properties of the product should improve. Improved phys-
ical characteristics will cause agricultural residue-based
fungal composites to be suitable for numerous applications
that presently use fossil-fuel based materials. Delivered by
Greg Holtchitinous material using fungal fruiting bodies and product made
Abbreviations IP : 199.133.133.66
Thu, 29 Nov 2012
CGB: Cotton Gin Byproducts
Disclaimer
Mention of product or trade names does not constitute
an endorsement by the USDA-ARS over other compara-
ble products. Products or trade names are listed for ref-
erence only. USDA is an equal opportunity provider and
employer.
Acknowledgment: This project was a collaborative
effort involving USDA-ARS in Lubbock, TX, and
Ecovative Design, LLC of Green Island, NY, under a
Cooperative Research and Development Agreement (#58-
3K95-0-1391). The authors would like to thank Philip
Pearson, Bill Turner, Jeff Turner, Jimmie Castro, Gary
Schlabs, Clinton Cox, and Chris Arinder for their work
in preparation of this manuscript, processing the cotton
byproducts, and making the blends used in this study.
References
1. http://en.wikipedia.org/wiki/Polystyrene. Verified, June (2011).
2. http://www.sciencedaily.com/releases/2009/08/090819234651.htm.
August 20 (2009). Verified June 2011.
3. K. G. Satyanarayana, G. G. C. Arizaga, and F. Wypych, Prog. Polym.
Sci. 34, 983 (2009).
4. X. Lu, M. Q. Zhang, M. Z. Rong, D. L. Yue, and G. C. Yang,
Compos. Sci. Technol. 64, 1301 (2004).
5. X. Zhou, F. Zheng, H. Li, and C. Lu, Energy and Buildings 42, 1070
(2010).
J. Biobased Mater. Bioenergy 6, 431–439, 2012
6. S. S. Ahankari, A. K. Mohanty, and M. Misra, Compos. Sci. Technol.
71, 653 (2011).
7. H. Gialamas, K. G. Zinoviadou, C. G. Biliaderis, and
K. P. Koutsoumanis, Food Research International 43, 2402 (2010).
8. Y. Makino and T. Hirata, Postharvest Biology and Technology
10, 247 (1997).
9. M. Rinaudo, Prog. Polym. Sci. 31, 603 (2006).
10. K. V. H. Prashanth and R. N. Tharanathan, Trends in Food Science
and Technology 18, 117 (2007).
11. I. O. Fasids and U. U. Ekuere, Food Chem. 48, 255 (1993).
12. I. O. Fasids, Food Chem. 55, 161 (1996).
13. A. Philippoussis, R. Diamantopoulou, and C. Israilides, Int.
Biodeterior. Biodegrad. 59, 216 (2007).
14. A. Medina, M. Vassileva, F. Caravaca, A. Roldán, and R. Azcón,
Chemosphere 56, 499 (2004).
15. J. B. Sutherland, Mycologia 76, 369 (1984).
16. Y. Li and K. L. Pickering, Compos. Sci. Technol. 69, 1265 (2009).
17. Y. Li, K. L. Pickering, and R. L. Farrell, Industrial Crops and
RESEARCH ARTICLE
Products 29, 420 (2009).
18. E. Bayer, G. McIntyre, and B. L. Swersey, Method for producing
grown materials and products made thereby, U.S. Patent Application
US 2008/0145577 A1, June (2008).
Ingenta
19. E. Bayerto:and G. McIntyre, Method for producing rapidly renewable
thereby. U.S. Patent Application US2009/0307969 A1, December
(2009).
23:20:57
20. ASTM C165-07, Standard Test Method for Measuring Compressive
Properties fo Thermal Insulations, ASTM International (2007).
21. ASTM C203-05a, Standard Test Methods for Breaking Load
and Flexural Properties of Block-Type Thermal Insulation, ASTM
International (2005).
22. ASTM C303-10, Standard Test Method for Dimensions and Den-
sity of Performed Block and Board-Type Thermal Insulation, ASTM
International (2010).
23. ASTM C481-99, Standard Test Method for Laboratory Aging of
Sandwich Constructions, ASTM International (2005).
24. ASTM C1134-90, Standard Test Method for Water Retention of
Rigid Thermal Insulations Following Partial Immersion, ASTM
International (2007).
25. ASTM E1354-11, Standard Test Method for Heat and Visible Smoke
Release Rates for Materials and Products Using and Oxygen Con-
sumption Calorimeter, ASTM International (2011).
26. Therm Test, Inc., TPS 500 thermal conductivity instrument infor-
mation booklet. http://www.thermtest.com/content/237301. Verified,
June (2011).
27. G. A. Holt, G. L. Barker, R. V. Baker, and A. D. Brashears, Trans.
ASAE 43, 1393 (2000).
28. Typical Physical Properties of Expanded Polystyrene, Undated http://
www.universalconstructionfoam.com/downloads/eps-data-sheet.pdf,
Verified, September (2011).
29. EPS Geofoam Data Sheet, Undated. http://www.
universalconstructionfoam.com/downloads/geofoam-eps-data-sheet.
pdf. Verified, September (2011).
30. Properties, Performance and Design Fundamentals of Expanded
Polystyrene Packaging 2000, http://www.epspackaging.org/images/
stories/Physical_Prop_Tech_Bull.pdf, Verified, September (2011).
31. R. N. Walters, S. M. Haackett, and R. E. Lyon, Undated, Heats
of combustion of high temperature polymers, www.fire.tc.faa.gov/
pdf/chemlab/hoc.pdf Verified, September (2011).
32. F. P. Incropera, D. P. Dewitt, T. L. Bergman, and A. S. Lavine,
Fundamentals of Heat and Mass Transfer, 6th edn., John Wiley &
Sons, Inc., Hoboken (2007).
Received: 28 July 2011. Accepted: 2 April 2012.
439