Mineral Composition, Growth Performance

and Yield of Pleurotus ostreatus on Rice

Straw-Based Substrate Enriched with Natural
Calcium Sources

Arianne V. Julian, Michael R. Umagat, and Renato G. Reyes

Keywords

Calcium fortification Pleurotus ostreatus Mushroom production Agricultural lime
Eggshell Oyster shell

1 Introduction et al. 2005). Cultivation of Pleurotus ostreatus is widespread

Calcium plays a vital role in maintaining optimum health
particularly the sustenance of bones and teeth; hence, ade-
quate daily intake is necessary. Most individuals can acquire
calcium by eating a variety of foods such as milk and other
dairy products; green, leafy vegetables; seafood, nuts, and
dried beans (Pravina et al. 2013). In this study, inedible
sources were used including agricultural lime, eggshells and
oyster shells which contain calcium carbonate as a main
component (Choi et al. 2011). Accordingly, bioconversion
of waste materials such as eggshells and oyster shells as
sources for Ca supplements provides an excellent method to
use renewable resources. Within the same trend, several
studies demonstrated the ability of mushrooms for nutrient
uptake from substrate (Ogundele et al. 2014). Substrate
supplementation is a common practice not only to improve
the nutritional values but also to enhance the yield (Moda
in different parts of the world which can be attributed to its
low cost production and its nutritional and medicinal prop-
erties (Kortei 2011). These mushrooms are an excellent
source of proteins, minerals such as phosphorus, iron,
potassium and sodium, and vitamin C, B complex such as
thiamine, riboflavin, folic acid and niacin (Çağlarırmak
2007). However, the total calcium content in mushrooms
remains low compared to vegetables (Kawai et al. 1990).
The present study evaluated the nutritive values, growth and
yield potential of P. ostreatus after cultivation on substrates
enriched with calcium using eggshells, oyster shells and
agricultural lime, and determined the concentration that
could be used for a successful production of
calcium-enriched mushrooms.
2 Materials and Methods

I. Source and Revival of Pure Culture

Mycelial discs from the pure culture of P. ostreatus were

A. V. Julian (&)
Department of Biological Sciences, College of Arts and Sciences,
Central Luzon State University, Nueva Ecija, Philippines
e-mail: ariannejulian@gmail.com
M. R. Umagat
Laboratory of Industrial Biotechnology and Bio-Industry,
Central Luzon State University, Nueva Ecija, Philippines
e-mail: michael.umagat@clsu.edu.ph
R. G. Reyes
Center for Tropical Mushroom Research and Development,
Central Luzon State University, Nueva Ecija, Philippines
e-mail: renato.reyes@clsu.edu.ph
aseptically transferred into culture plates and incubated for
seven days at room temperature to allow mycelial growth.
II. Ca Sources Collection and Powder Preparation
Eggshells and oyster shells were washed with water and cut
into small pieces, then sun-dried for 24 h and placed in an
oven for 30 min to completely dry the samples. Dried egg-
shells and oyster shells were powderized using a blender and
sifted using a metal sieve.

© Springer International Publishing AG 2018 1573

A. Kallel et al. (eds.), Recent Advances in Environmental Science from the Euro-Mediterranean and Surrounding Regions,
Advances in Science, Technology & Innovation, https://doi.org/10.1007/978-3-319-70548-4_457
1574 A. V. Julian et al.

III. Substrate Formulation, Preparation and Table 1 Influence of varying concentrations of Ca sources on the
Enrichment yield and biological efficiency of P. ostreatus
Calcium Concentration Total yield (g) Biological
sources (%) efficiency (%)
Seven parts of composted rice straw were combined with
three parts of sawdust and one part of rice bran. The mixture Agricultural 0 150.60 ± 4.22b 28.69 ± 0.93b
lime
was divided and supplemented with different concentrations 2 179.20 ± 14.55 ab
33.52 ± 2.98ab
(2, 4, 6, 8 and 10%) of the Ca sources and the unsupple- 4 177.40 ± 26.58ab 33.70 ± 5.18ab
mented mixture served as control. Enriched substrates were 6 165.80 ± 19.66 ab
31.27 ± 3.50ab
compressed in polypropylene bags and sterilized for 1 h. 8 191.80 ± 19.10a 35.57 ± 2.99a
The bags were allowed to cool down and each bag was 10 191.20 ± 18.14 a
35.63 ± 3.75a
aseptically inoculated with grain spawn. Initial weights of Eggshell 0 150.60 ± 4.22 b
28.69 ± 0.93b
the bags were obtained and were incubated at 30 °C tem- powder
2 170.80 ± 28.66ab 31.88 ± 5.28ab
perature with 80–90% relative humidity until mushroom
4 169.20 ± 19.07 ab
31.51 ± 3.43ab
formation.
6 197.60 ± 19.05 a
37.43 ± 5.54a
8 154.60 ± 17.39b 29.89 ± 3.52b
IV. Harvest and Assessment of Biological Efficiency
10 143.20 ± 10.20 b
27.26 ± 2.51b
The duration of the period from inoculation to full mycelial Oyster shell 0 150.60 ± 4.22a 28.69 ± 0.93a
powder
colonization, primordial formation and fruiting body devel- 2 172.40 ± 18.23 a
32.82 ± 3.58a
opment were recorded. The fruiting bodies of P. ostreatus 4 173.20 ± 25.17 a
33.04 ± 4.42a
were then harvested, weighed and air-dried until the 3rd 6 168.00 ± 8.22a 32.07 ± 1.50a
flush. Yield and biological efficiency (BE) were recorded. 8 153.60 ± 20.38 a
29.18 ± 3.86a
10 168.40 ± 37.96 a
31.98 ± 7.43a
V. Determination of Mineral Contents in Fruiting The displayed values are means and SD. Treatment means in each Ca source
Bodies with different letter of superscript in each column are significantly different
from each other at 5% level of significance using LSD
Dried fruiting bodies of P. ostreatus were ground using a
blender and analyzed for elemental composition using
Thermo EDX system installed in Hitachi SU1510 scanning
electron microscope. Other elements such as Mg, P, K, Na,
Si, Cl, and S were also determined.

3 Results and Discussion

The findings of this study revealed that the use of natural Ca

sources as substrate additives generally increased the yield,
biological efficiency and Ca content of P. ostreatus. The
duration of days from inoculation to mycelial colonization,
primordial initiation and first flush were slightly prolonged.
Mushroom grown in substrate with 8 and 10% agricultural
lime produced the highest yields with 191.80 and 191.20 g,
respectively. In eggshell powder, the superior total yield was
recorded in 6% with 197.60 g while in oyster shell
powder-supplemented mushrooms, the highest yields were
recorded in 2 and 4% with 172.30 and 173.20 g, respectively
(Table 1).
In agricultural lime-supplemented substrate, the biologi-
cal efficiency (BE) was highest in 8% and 10% with 35.57
and 35.63%, respectively. Meanwhile, the highest BE in
eggshell powder-supplemented substrate was noted at 6%
with 37.43%. Similarly, oyster shell powder enrichment
Fig. 1 Average content of Ca absorbed by P. ostreatus in every
concentration of different Ca sources
increased the BE of P. ostreatus ranging from 29.18 to
33.82% but not statistically different from the control
(Table 1). The optimum Ca contents in P. ostreatus culti-
vated in rice straw-based substrate were recorded in 6%
agricultural lime (9.06%), 8% agricultural lime (9.66%) and
10% eggshell powder (9.51%). While the addition of these
sources both resulted in Ca absorption of P. ostreatus, oyster
shell powder supplementation at all levels showed no Ca
absorption efficacy (Fig. 1).
On the other hand, supplementation of all Ca sources
reduced K and P contents; however, the levels of Mg, Na, Si,
Cl and S were generally increased. These results suggest that
Mineral Composition, Growth Performance and Yield …
the nutritional value of P. ostreatus mushroom can be sig-
nificantly improved with the addition of Ca sources.
4 Conclusion
Varying the Ca concentrations from different sources
affected the yield and mineral absorption efficacy of
P. ostreatus mushroom. Based on the findings of the
present study, it can be concluded that agricultural lime
and eggshell powder (up to 10%), but not oyster shell
powder, generally increased the yield and biological
efficiency. This growth improvement may signify an
association with increased Ca content (less than 10%)
recorded in the fruiting bodies of mushroom. Concen-
trations of other minerals such as Mg, Na, Si, Cl and S
were generally increased but decreased K and P levels in
the supplemented mushrooms were lower than those in
the control. This study suggests that using Ca-rich bio-
logical wastes may improve the Ca concentration in
P. ostreatus, but may also induce diverse effects on the
absorption of other essential minerals.
1575
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