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Volume 21(1), 150- 157, 2017 JOURNAL of Horticulture, Forestry and Biotechnology www.journal-hfb.usab-tm.ro
150
Study on chemical composition of Agaricus blazei Murrill mushroom, produced on different substrates Rózsa S.1*, Măniuțiu D.N.1, Gocan Tincuța-Marta1, Sima Rodica1, Rózsa Melinda2
1University of Agricultural Sciences and Veterinary Medicine, Faculty of Horticulture 3-5 Mănăștur St.,
400372, Cluj-Napoca, România; 2 SC Ciupercăria SRL, 332 Aghireșu-Fabrici, 407010, Cluj County,
România, www.ciupercaria.com *Corresponding author. Email: [email protected] Abstract All edible mushrooms are high in vitamin B plus other vitamins such as vitamin C and ergosterol. Bioactive compounds from edible mushrooms have become new products for health therapy, especially anti-cancer therapies. Almond mushroom - Agaricus blazei Murrill was discovered and popularised as late as the 20th century. However, it has been known for its exceptional properties in the places of its origin for a long time. Studies have been conducted worldwide for several decades, aiming at the precise determination of these properties and their applicability in medicine. This study was carried out to evaluate the effect of four growing substrates of Agaricus blazei Murrill mushroom on the chemical composition of mushrooms.
Key words compost, Agaricus blazei Murrill, vitamin B, vitamin C, ergosterol
The fruiting bodies of Agaricus blazei Murrill contain
89-91% water, which is in general less than that of A.
bisporus. Almost 48% of total dry matter consists of
crude protein and 18% of carbohydrates, but the lipid
content is only 0.5% [6]. The fruiting bodies of
Agaricus blazei Murrill contain high levels of valuable
minerals, e.g. potassium, phosphorus, calcium,
magnesium and zinc. Nevertheless, a minute amount of
cadmium was also detectable [6].
Bioactive compounds of mushrooms can be isolated
from their fruiting bodies, or culture extraction from
pure culture of mycelia [1]. Agaricus blazei Murrill has
been reported to produce various bioactive compounds
that have potential to treat many diseases [5]. This
mushroom has been used as a medicinal food for the
prevention of cancer, diabetes, hyperlipidaemia,
arteriosclerosis, and chronic hepatitis, and is known to
stimulate the immune system [20].
Several categories of molecules are involved in
beneficial effects of Agaricus blazei Murrill and most
of them are common to the entire fungal kingdom [10].
Some compounds (such as ergosterol and b-glucans)
are considered as biochemical markers for the kingdom
and are ubiquitous. β-Glucans are cell wall constituents
that can be found in many fungi [10].
Agaricus blazei Murrill is a well-known medicinal
mushroom used in many countries, and thus
consumption of this mushroom is used as an alternative
way to cure diseases. Various pharmaceutical activities
have been found associated with Agaricus blazei
Murrill and researches to reveal the function of
bioactive compounds are extensive. Recent studies
have been performed in vitro and in vivo to confirm the
mushrooms therapeutic properties [5]. Identification of
(novel) immunomodulating bioactive compounds from
the mushroom may also help in new treatments for
patients suffering from cancer and immunodeficiency
[13].
Almond mushroom is a saprophytic species, growing
well on soils rich in ligno-cellulose residue [19]. As it
was reported by Stamets [19], Wasser [21] and Dias
[4], natural habitats for this species include mixed
forests, first of all their margins, as well as fields in
upland and mountainous areas of Brazil and Peru.
Almond mushroom is the so-called secondary
saprophyte, developing on partially processed
substrate, in which microorganisms reduced complex
ligno-cellulose compounds [2]. Numerous authors have
shown that due to the similar life cycle in the
cultivation of almond mushroom technologies
developed for white button mushroom may be applied.
However, almond mushroom requires high temperature
and high humidity as well as access to light to form
fruiting bodies [2,3,4,11,18]. In Brazil, due to the
advantageous climatic conditions this species is
frequently grown outdoors; however, in other countries
- mainly due to its high temperature requirements -
such cultivation system is risky and may only be
successful during very warm summers [9,18,19].
The results of Agaricus cultivation to a considerable
degree are determined by the composition of the
substrate. Unfortunately, there is a limited body of data
concerning growing media for almond mushroom
cultivation. Frequently producers, particularly in Brazil
and Japan, use substrate with the composition
developed for white button mushroom [11]. However,
151
both species may respond differently to an identical
substrate composition. For this reason, it is essential to
develop a substrate meeting requirements of almond
mushroom [22]. The primary components in such a
substrate are most frequently locally available
materials, subjected to composting, e.g. agricultural
waste rich in ligno-cellulose complexes, i.e. straw,
cotton burrs, grasses, sawdust, enriched with animal
manure, poultry dung, wheat or rice bran and calcium
[7,8,9,12,14,17].
Material and Method
Given that in the experience we made, the culture
substrate presents 4 graduations (a1-classic compost,
a2- synthetic compost, a3-mixed compost, a4-original
compost) to achieve directed composting, were
conducted 4 identical tanks to control, perform and
record optimal environmental conditions necessary for
the composting and pasteurization process. (figure no.
1-6).
For the experiments, we used four recipes of compost, presented in table no. 1
7
4
6 5
1
2 3
8
9
1
0
Module a1
11
7
4
6 5
1
2 3
8
9
1
0
Module a2
7
4
6 5
1
2 3
8
9
1
0
Module a3
7
4
6 5
1
2 3
8
9
1
0
Module a4
Figure no. 1. Sketch for the composting facility - 4 modules:
1 - tank with capacity of 1 m3 for compost components; 2 – rack for compost; 3 - tank heating system for composting;
4 - tank for the collection and recirculation the water excess; 5 - heating elements for wetting water (purine); 6 –
water/purine recirculation pump; 7 - recirculation pipes for wetting water/ purine; 8 - compost discharge door; 9 - air
flow control valve for aerobic composting; 10 - air inlet pipe to aerobic composting from the compressor; 11 -
ground level.
Figure 2. Composting facility - 4
modules
Source: original photo
Figure 3. Tank for the collection and
recirculation of the water excess
Source: original photo
Figure 4. Compressor for the air flow
Source: original photo
152
Table 1
Recipes for compost used in experience
Type of compost
Components Quantity for 1 tone of
compost
C1 - Classical Horse manure (horse manure and wheat bedding straw 70-75%)
Gypsum (calcium sulphate)
Superphosphate
Ammonium sulphate
500 kg
25 kg
7 kg
7 kg
C2 - Synthetic Wheat straw
Poultry litter
Gypsum (calcium sulphate)
Urea
350 kg
150 kg
20 kg
7 kg
C3 - Mixt Horse manure (horse manure and wheat bedding straw 70-75%)
Poultry litter
Wheat straw
Gypsum (calcium sulphate)
Urea
250 kg
100 kg
150 kg
24 kg
2 kg
C4 - Original shredded reed
Horse manure (horse manure and wheat bedding straw 70-75%)
Poultry litter
Gypsum (calcium sulphate)
Urea
100 kg
200 kg
150 kg
24 kg
2 kg
Each type of compost was prepared in a composting
tank, respecting the proportions of raw and auxiliary
materials.
The compost pasteurization was achieved by raising
the temperature of compost at 58-60 °C for a period of
about 8 hours, then lowering the temperature of the
compost at 50°C by mixing fresh air and continued
cooling to 45°C. The temperature of 45°C was
maintained until the ammonia content of the compost
has not fallen below 0.05% and the pH has stabilized in
the range of 7.3-7.5. [15,16]. In experience, we used
protein additives as follows: A1 – without protein
additives, A2 - 3% wheat bran and A3 - Corn flour 3%.
Results and Discussions
Table no. 2 presents the effect of compost and protein
additives (A x C) on the dry matter of mushrooms
during year 2015 and 2016 of experience.
The maximum value of total dry matter (9.43%) and
soluble dry matter (8.50%) was recorded by mushroom
harvested from C3 A2 combination, the minimum
value of total dry matter was recorded by C4 A1
combination (7.13%) and the minimum value of
soluble dry matter was recorded by C4 A3 combination
(7.07%).
Table no. 3 presents the effect of the interaction
between protein additives A and compost C to the
soluble dry matter content of mushrooms, during year
2015 and 2016 of experience.
Figure 5. Tank for composting, internal view.
Source: original photo
Figure 6. Compost components pre-soaking.
Source: original photo
153
Table 2
Effect of compost and protein additives (A x C) on the dry matter of mushrooms during two years of experience
Experimental
factors
Content of:
Total dry matter % Soluble dry matter % Insoluble dry matter %
Variant Combi
nation 2015 2016 media 2015 2016 media 2015 2016 media
V1 C1 A1 9.03 8.23 8.63 7.73 7.07 7.40 1.30 1.16 1.23
V2 C1 A2 9.43 8.70 9.07 8.80 8.13 8.47 0.63 0.57 0.60
V3 C1 A3 9.73 8.87 9.30 8.07 7.40 7.73 1.66 1.47 1.57
V4 C2 A1 8.20 7.57 7.87 8.17 7.30 7.75 0.13 0.27 0.12
V5 C2 A2 9.10 8.50 8.80 8.50 7.77 8.13 0.60 0.73 0.67
V6 C2 A3 8.70 7.77 8.23 8.33 7.73 8.03 0.37 0.04 0.20
V7 C3 A1 9.40 8.60 9.00 7.93 7.27 7.60 1.47 1.33 1.40
V8 C3 A2 9.77 9.10 9.43 8.83 8.17 8.50 0.94 0.93 0.93
V9 C3 A3 8.47 7.80 8.13 7.43 6.77 7.10 1.04 1.03 1.03
V10 C4 A1 7.60 6.83 7.13 7.37 6.67 7.10 0.23 0.16 0.03
V11 C4 A2 8.37 7.83 8.10 7.63 6.87 7.25 0.74 0.96 0.85
V12 C4 A3 7.70 7.17 7.43 7.57 6.57 7.07 0.13 0.60 0.36
Table 3
The effect of the interaction between protein additives A and compost C
to the soluble dry matter content of mushrooms
Experimental
variant Combination
Soluble dry
matter SDM% Significance*
V6 A2 C2 8.80 A
V7 A2 C3 8.50 B
V5 A2 C1 8.47 B
V11 A3 C3 8.13 C
V8 A2 C4 8.10 C
V10 A3 C2 8.03 C
V2 A1 C2 7.87 CD
V9 A3 C1 7.73 D
V3 A1 C3 7.60 E
V12 A3 C4 7.43 F
V1 A1 C1 7.40 F
V4 A1 C4 7.10 G
DS 0.16-0.18
*Values marked with different letters are significant
Taking into account the combined influence of protein
additives and the compost on the soluble dry matter
content of mushrooms, the best value was recorded by
A2 C2 combination with 8.80% SDM, followed by A2
C3 combination with 8.50% SDM. On the last places
was A1 C4 combination with 7.10% SDM.
Table no. 4 presents the results obtained for B1 vitamin
(thiamine), in μg, during year 2015 and 2016 of
experience.
154
Table 4
B1 vitamin (thiamine), in μg, during year 2015 and 2016 of experience
Experimental factors B1 vitamin μg B1 vitamin processing value with Duncan Test
(2015-2016) μg
Variant Combination 2015 2016 Variant Combination Value Signification
V1 C1 A1 1053 1007 V9 A3 C1 1151.67 A
V2 C1 A2 1123 1077 V5 A2 C1 1100.00 B
V3 C1 A3 1180 1123 V1 A1 C1 1030.00 C
V4 C2 A1 923 877 V10 A3 C2 956.67 D
V5 C2 A2 963 917 V6 A2 C2 940.00 D
V6 C2 A3 980 933 V2 A1 C2 900.00 E
V7 C3 A1 600 547 V11 A3 C3 635.00 F
V8 C3 A2 640 593 V7 A2 C3 616.67 F
V9 C3 A3 667 603 V3 A1 C3 573.33 G
V10 C4 A1 403 360 V12 A3 C4 423.33 H
V11 C4 A2 427 363 V8 A2 C4 395.00 HI
V12 C4 A3 447 400 V4 A1 C4 381.67 I
Mean of
experience 783.83 733.33 DS 35.68----41.15
Following the values from table no. 4, we can see that
the maximum content of B1 vitamin was recorded by
C1 compost (1007-1180μg), followed by C2 compost
(877-980μg) harvested mushrooms.
On the last place was mushrooms harvested from the
C4 compost (363-447μg). As we can see the protein
additives did not influenced the B1 vitamin content so
much as the type of compost. On the first place is
situated A3 protein additives, followed by A2 and A1
protein additives.
Table no. 5 presents the results obtained for B2 vitamin
(riboflavin), in μg, during year 2015 and 2016 of
experience.
Table 5
B2 vitamin (riboflavin), in μg, during year 2015 and 2016 of experience
Experimental factors B2 vitamin μg B2 vitamin processing value with Duncan Test
(2015-2016) μg
Variant Combination 2015 2016 Variant Combination Value Signification
V1 C1 A1 5300 4667 V7 A2 C3 5983.33 A
V2 C1 A2 5667 5233 V9 A3 C1 5616.67 AB
V3 C1 A3 5800 5433 V5 A2 C1 5433.33 ABC
V4 C2 A1 4033 3600 V11 A3 C3 5350.00 ABCD
V5 C2 A2 4533 4100 V1 A1 C1 4983.33 ABCD
V6 C2 A3 4767 4333 V3 A1 C3 4800.00 BCDE
V7 C3 A1 5000 4600 V10 A3 C2 4550.00 CDEF
V8 C3 A2 5300 4667 V6 A2 C2 4316.67 DEFG
V9 C3 A3 5533 5167 V2 A1 C2 3816.67 EFGH
V10 C4 A1 3400 2967 V12 A3 C4 3533.33 FGH
V11 C4 A2 3700 3200 V8 A2 C4 3450.00 GH
V12 C4 A3 3767 3300 V4 A1 C4 3183.33 H
Mean of
experience 4733.3 4272.2 DS 944.83----1089.71
Following the values from table no. 5, we can see that
the maximum content of B2 vitamin was recorded in
2015 by C1 compost with A3 protein additives
(5800μg), followed by C1 compost with A2 protein
additives (5667μg) harvested mushrooms. On the last
place was mushrooms harvested from the C4 compost
with A1 protein additives (3400μg). In 2016 that the
maximum content of B2 vitamin was recorded by C1
compost with A3 protein additives (5443μg) and the
minimum value was recorded also by C4 compost with
A1 protein additives (2967μg).
As we can see the combined factors, protein additives
and compost, A2 C3 combination recorded the
maximum value (5983.33μg), followed by A3 C1
combination. On the last place is situated the A1 C4
combination (3183.33μg).
Table no. 6 presents the results obtained for B9 vitamin
(folic acid), in μg, during year 2015 and 2016 of
experience.
155
Table 6
B9 vitamin (folic acid), in μg, during year 2015 and 2016 of experience
Experimental factors B9 vitamin μg B9 vitamin processing value with Duncan Test
(2015-2016) μg
Variant Combination 2015 2016 Variant Combination Value Signification
V1 C1 A1 580 567 V1 A3 C3 671.67 A
V2 C1 A2 610 587 V9 A3 C1 631.67 B
V3 C1 A3 643 620 V7 A2 C3 631.67 B
V4 C2 A1 443 427 V3 A1 C3 618.33 B
V5 C2 A2 467 450 V5 A2 C1 598.33 C
V6 C2 A3 483 467 V1 A1 C1 579.33 D
V7 C3 A1 643 593 V10 A3 C2 475.00 E
V8 C3 A2 673 590 V6 A2 C2 458.33 F
V9 C3 A3 687 657 V2 A1 C2 435.00 G
V10 C4 A1 290 277 V12 A3 C4 313.33 H
V11 C4 A2 303 280 V8 A2 C4 291.67 I
V12 C4 A3 323 303 V4 A1 C4 283.33 I
Mean of
experience 512.08 484.8 DS 13.77----15.88
Following the values from table no. 6, we can see that
the maximum content of B9 vitamin was recorded in
2015 by C3 compost with A3 protein additives
(687μg), followed by C3 compost with A2 protein
additives (673μg) harvested mushrooms. On the last
place was mushrooms harvested from the C4 compost
with A1 protein additives (323μg). In 2016 that the
maximum content of B9 vitamin was recorded by C3
compost with A3 protein additives (657μg) and the
minimum value was recorded also by C4 compost with
A1 protein additives (277μg).
As we can see the combined factors, protein additives
and compost, A3 C3 recorded the maximum value
(671.67μg), followed by A3 C1 combination. On the
last place is situated the A1 C4 combination
(283.33μg).
Table no. 7 presents the results obtained for B12
vitamin (cobalamin), in μg, during year 2015 and 2016
of experience.
Table 7
B12 vitamin (cobalamin), in μg, during year 2015 and 2016 of experience
Experimental factors B12 vitamin μg B12 vitamin processing value with Duncan Test
(2015-2016) μg
Variant Combination 2015 2016 Variant Combination Value Signification
V1 C1 A1 527 510 V11 A3 C3 906.67 A
V2 C1 A2 540 523 V7 A2 C3 866.67 B
V3 C1 A3 560 557 V3 A1 C3 828.33 C
V4 C2 A1 727 680 V10 A3 C2 761.67 D
V5 C2 A2 747 710 V6 A2 C2 728.33 E
V6 C2 A3 767 757 V2 A1 C2 703.33 E
V7 C3 A1 843 813 V12 A3 C4 573.33 F
V8 C3 A2 877 857 V9 A3 C1 558.33 FG
V9 C3 A3 913 900 V8 A2 C4 538.33 GH
V10 C4 A1 487 440 V5 A2 C1 531.67 GH
V11 C4 A2 550 527 V1 A1 C1 518.33 H
V12 C4 A3 583 563 V4 A1 C4 463.33 I
Mean of
experience 676,75 653.08 DS 32.40----37.37
Following the values from table no. 7, we can see that
the maximum content of B12 vitamin was recorded in
2015 by C3 compost with A3 protein additives
(913μg), followed by C3 compost with A2 protein
additives (877μg) harvested mushrooms. On the last
place was mushrooms harvested from the C4 compost
with A1 protein additives (487μg). In 2016 that the
maximum content of B12 vitamin was recorded by C3
compost with A3 protein additives (900μg) and the
minimum value was recorded also by C4 compost with
A1 protein additives (440μg).
As we can see the combined factors, protein additives
and compost, A3 C3 recorded the maximum value
(906.67μg), followed by A2 C3 combination. On the
156
last place is situated the A1 C4 combination
(463.33μg).
Table no. 8, presents the unilateral influence of the
compost on the mushroom C vitamin contents, in mg,
during year 2015 and 2016 of experience
Table 8
The unilateral influence of the compost on the mushroom C vitamin contents
C
O
M
P
O
S
T
Year
2015 2016
C vitamin
Ascorbic acid
C vitamin
Ascorbic acid
Absolute
value
Relative
value
± D
%
Signification of
difference
Absolute
value
Relative
value
± D
%
Signification of
difference
C1 22.56 120.8 3.89 ** 16.89 111.4 1.72 *
C2 15.89 85.1 -2.78 0 13.0 85.7 -2.17 0
C3 21.00 112.5 2.33 * 18.22 120.1 3.06 **
C4 15.22 81.5 -3.44 00 12.56 82.8 -2.61 00
Avg. 18.67 100.0 0,00 Mt. 15.17 100.0 0.00 Mt
DL/LSD 5% 1.88 1.58
DL/LSD 1% 2.84 2.39
DL/LSD 0,1% 4.56 3.83
Following the values from table no. 8, we can see that
mushroom harvested from C1 and C3 composts,
presented significant C vitamin differences from the
average.
Conclusions
Following the results presented of the above, the
mushrooms harvested on C3 compost with A2 protein
additives presents the highest total and soluble dry
matter content. If we want to obtain mushrooms with a
high level of B1 and B2 vitamin, we can use the C1
compost with A3 protein additives.
For obtaining mushrooms with a high level of B9 or
B12 vitamin, we recommended to use the C3 compost
with A3 protein additives. For obtaining mushrooms
with a highest level of C vitamin, we can use C1 or C3
composts.
The A3 protein additive is more efficient in increasing
the vitamin content of harvested mushrooms.
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