Influence of the Culture Substrate on the Agaricus blazei Murrill Mushrooms Vitamins Content

The vitamin content of cultivated mushrooms differs from one species to another, depending on their stage of development, the nutrient substrate used to produce them, and the microclimate in the culture space. Agaricus blazei Murrill is one of the most popular cultivated medicinal mushrooms, with scientifically proven therapeutic properties. Considering that the Agaricus spp. mushrooms culture substrate can be produced using various raw materials, in this paper we have studied the influence of the culture substrate using four types of substrate with different protein additions on the vitamin content of mushrooms. The food qualities of the Agaricus blazei Murrill mushrooms, evaluated by the chemical composition, generally revealed the product obtained on the classic compost, improved with the addition of proteinaceous of corn flour. Mushrooms harvested on this substrate have the highest levels of B1 (1151 μg 100 g−1 dm), B9 (671 μg 100 g−1 dm), B12 (906 μg 100 g−1 dm), PP (55.33 μg 100 g−1 dm), and C vitamins (21.67 μg 100 g−1 dm). The content of ergosterol, as a precursor of D2 vitamin, has higher values in the product obtained on the classic compost, with the addition of wheat bran (90.17 mg 100 g−1 dm) and the addition of corn flour (94 mg 100 g−1 dm).


Introduction
The food value of a product depends mainly on two factors, as follows: The digestibility of the compounds that make up its composition and the chemical composition [1]. The chemical analyses of common edible mushrooms showed that, on the basis of fresh substance, the moisture content is 89-91%, the ash is 0.97-1.26%, the protein is 1.78-4.94%, the fat is 0.1-0.65%, crude fiber is 0.09-1.67%, carbohydrates are 2.3-6.28%, and the energy value is 24.4-34.4 Kcal. Glucose is present in very small amounts and, like any other vegetable, the mushrooms contain very small amounts of fat. Their caloric value is rather low compared to other foods, so they can be recommended in slimming diets [2,3].
The vitamin content shows that among all plant products, mushrooms are real sources of vitamins, especially those of complex B (thiamine, biotin, nicotinic acid, pantothenic acid) and vitamin D, which is no longer found in any other plant, being specific to fish lard and meat [4]. In addition, in cultivated mushrooms there are significant amounts of vitamin A, A 1 , and vitamin C (ascorbic acid). With regard

Ergosterol Content of Agaricus blazei Murrill Mushrooms (mg 100g −1 dm)
The content of ergosterol (mg 100g −1 dm) in Agaricus blazei Murrill mushrooms varied between 75.00 and 95. 67   In both experimental years, the maximum value was recorded in the mushrooms harvested on the classical compost with a protein admixture of corn flour (V3), with a maximum of 95.67 mg 100g −1 dm in 2017 and 92.33 mg 100g −1 dm in 2018.
The lowest content of ergosterol was recorded in mushrooms harvested from cane compost without additional protein supplement (V10), with 75.00 mg 100g −1 dm in 2017 and 71.00 mg 100g −1 dm in 2018. In both experimental years, the maximum value was recorded in the mushrooms harvested on the classical compost with a protein admixture of corn flour (V3), with a maximum of 95.67 mg 100 g −1 dm in 2017 and 92.33 mg 100 g −1 dm in 2018.
The lowest content of ergosterol was recorded in mushrooms harvested from cane compost without additional protein supplement (V10), with 75.00 mg 100 g −1 dm in 2017 and 71.00 mg 100 g −1 dm in 2018. The vitamin C content (mg 100 g −1 dm) in Agaricus blazei Murrill mushrooms varied between 12.33 and 24.33 mg 100 g −1 dm in 2017 and 9.67 and 20.33 mg in 2018 ( Figure 2).

Vitamin C Content of Agaricus blazei Murrill Mushrooms (mg 100g −1 dm)
The vitamin C content (mg 100g −1 dm) in Agaricus blazei Murrill mushrooms varied between 12.33 and 24.33 mg 100g −1 dm in 2017 and 9.67 and 20.33 mg in 2018. (Figure 2). In 2017, the maximum value was recorded for the mushrooms harvested on the classical compost with added corn flour protein (V3), with a maximum of 24.33mg 100g −1 dm, in 2018, the maximum value was recorded for mushrooms harvested from the composite compost with additional corn flour protein addition (V9).
Following the effect of interaction, protein × compost, in the years of experimentation on the vitamin C content (mg 100g −1 dm.) in Agaricus blazei Murrill mushrooms, it can be seen that the highest vitamin C content was recorded in the mushrooms harvested on the composite compost with the corn flour protein admixture (V9), with 21.67 mg 100g −1 dm, followed by the mushrooms harvested from the classic compost with corn flour (V3) with 21.50 mg 100g −1 dm of vitamin C.

B1 Vitamin (Thiamin) Content of Agaricus blazei Murrill Mushrooms (μg 100g −1 dm)
The content of vitamin B1 (μg 100g −1 dm) in Agaricus blazei Murrill mushrooms, ranged between 403 and 1180 μg 100g −1 dm in 2017 and between 360 and 1123 μg 100g −1 dm in 2018 ( Figure 3). In both experimental years, the maximum value was recorded in the mushrooms harvested on the classical compost with the corn flour protein admixture (V3).  In 2017, the maximum value was recorded for the mushrooms harvested on the classical compost with added corn flour protein (V3), with a maximum of 24.33 mg 100 g −1 dm, in 2018, the maximum value was recorded for mushrooms harvested from the composite compost with additional corn flour protein addition (V9).

V1
Following the effect of interaction, protein × compost, in the years of experimentation on the vitamin C content (mg 100 g −1 dm) in Agaricus blazei Murrill mushrooms, it can be seen that the highest vitamin C content was recorded in the mushrooms harvested on the composite compost with the corn flour protein admixture (V9), with 21.67 mg 100 g −1 dm, followed by the mushrooms harvested from the classic compost with corn flour (V3) with 21.50 mg 100 g −1 dm of vitamin C. The content of vitamin B 1 (µg 100 g −1 dm) in Agaricus blazei Murrill mushrooms, ranged between 403 and 1180 µg 100 g −1 dm in 2017 and between 360 and 1123 µg 100 g −1 dm in 2018 ( Figure 3). In both experimental years, the maximum value was recorded in the mushrooms harvested on the classical compost with the corn flour protein admixture (V3).
Analyzing the effect of the compost × additive interaction, on average, over the years of the experimental trial on the vitamin B 1 content (µg 100 g −1 dm), it can be noticed that the protein addition with corn flour (A3) increased the vitamin B 1 content of Agaricus blazei Murrill mushrooms on all four types of compost.
The highest values were obtained on the classical (C1) (1151 µg 100 g −1 dm) and the synthetic compost (C2) (956 µg 100 g −1 dm). Analyzing the effect of the compost × additive interaction, on average, over the years of the experimental trial on the vitamin B1 content (μg 100g −1 dm), it can be noticed that the protein addition with corn flour (A3) increased the vitamin B1 content of Agaricus blazei Murrill mushrooms on all four types of compost.

Vitamin B2 (Thiamin) Content of Agaricus blazei Murrill Mushrooms (μg 100g −1 dm)
The content of vitamin B2 (μg 100g −1 dm) in Agaricus blazei Murrill mushrooms ranged between 3400 and 5800 μg 100g −1 dm in 2017 and between 2967 and 5433 μg 100g −1 dm in 2018. (Figure 4). In both experimental years, the maximum value was recorded in the mushrooms harvested on the classical compost with the corn flour protein admixture (V3).  The content of vitamin B 2 (µg 100 g −1 dm) in Agaricus blazei Murrill mushrooms ranged between 3400 and 5800 µg 100 g −1 dm in 2017 and between 2967 and 5433 µg 100 g −1 dm in 2018 ( Figure 4). In both experimental years, the maximum value was recorded in the mushrooms harvested on the classical compost with the corn flour protein admixture (V3). Analyzing the effect of the compost × additive interaction, on average, over the years of the experimental trial on the vitamin B1 content (μg 100g −1 dm), it can be noticed that the protein addition with corn flour (A3) increased the vitamin B1 content of Agaricus blazei Murrill mushrooms on all four types of compost.

Vitamin B2 (Thiamin) Content of Agaricus blazei Murrill Mushrooms (μg 100g −1 dm)
The content of vitamin B2 (μg 100g −1 dm) in Agaricus blazei Murrill mushrooms ranged between 3400 and 5800 μg 100g −1 dm in 2017 and between 2967 and 5433 μg 100g −1 dm in 2018. (Figure 4). In both experimental years, the maximum value was recorded in the mushrooms harvested on the classical compost with the corn flour protein admixture (V3). Analyzing the effect of the compost × additive interaction in years of experimental trial on the vitamin B 2 content (µg 100 g −1 dm) of Agaricus blazei Murrill, it can be noticed that the protein addition of corn flour (A3) influenced the vitamin B 2 content of the mushrooms, the first place being the combination of corn flour protein admixture with classical compost (V3), with 5616 µg vitamin B 2 100 g −1 dm. The composite compost with wheat bran protein (V8), also influenced the vitamin B 2 content of the Agaricus blazei Murrill mushrooms (4983 µg 100 g −1 dm), but with statistically uninsured differences compared to the first variation.  Analyzing the effect of the compost × additive interaction in years of experimental trial on the vitamin B2 content (μg 100g −1 dm) of Agaricus blazei Murrill, it can be noticed that the protein addition of corn flour (A3) influenced the vitamin B2 content of the mushrooms, the first place being the combination of corn flour protein admixture with classical compost (V3), with 5616 μg vitamin B2 100g −1 dm. The composite compost with wheat bran protein (V8), also influenced the vitamin B2 content of the Agaricus blazei Murrill mushrooms (4983 μg 100g −1 dm), but with statistically uninsured differences compared to the first variation.

Vitamin B12 (Cobalamin) Content of Agaricus blazei Murrill Mushrooms (μg 100g −1 dm)
The content of vitamin B12 (μg 100g −1 dm) in Agaricus blazei Murrill mushrooms, ranged between 487 and 913 μg 100g −1 dm in 2017 and between 440 and 900 μg 100g −1 dm in 2018 ( Figure 6). In both experimental years the maximum value was recorded in the mushrooms harvested on the composite compost with the proteinaceous addition of corn flour (V9). The content of vitamin B 12 (µg 100 g −1 dm) in Agaricus blazei Murrill mushrooms, ranged between 487 and 913 µg 100 g −1 dm in 2017 and between 440 and 900 µg 100 g −1 dm in 2018 ( Figure 6). In both experimental years the maximum value was recorded in the mushrooms harvested on the composite compost with the proteinaceous addition of corn flour (V9).
The highest content of vitamin B 12 was found in mushrooms grown on mixed compost, especially that with the additional protein supplementation of corn flour (V9), followed closely, but with statistically ensured differences, by protein addition of bran wheat (V8). The highest content of vitamin B12 was found in mushrooms grown on mixed compost, especially that with the additional protein supplementation of corn flour (V9), followed closely, but with statistically ensured differences, by protein addition of bran wheat (V8).

PP Vitamin (B3-niacin) Content of Agaricus blazei Murrill Mushrooms (mg 100g −1 dm)
The content of the PP vitamin (mg 100g −1 dm) in Agaricus blazei Murrill mushrooms ranged between 30.33 and 56 mg 100g −1 dm in 2017 and between 29 and 54.67 mg 100g −1 dm in 2018, depending on the experimental compost variants used. (Figure 7). In both experimental years the maximum value was recorded in the mushrooms harvested on the composite compost with the proteinaceous addition of corn flour (V9).   The highest content of vitamin B12 was found in mushrooms grown on mixed compost, especially that with the additional protein supplementation of corn flour (V9), followed closely, but with statistically ensured differences, by protein addition of bran wheat (V8).

PP Vitamin (B3-niacin) Content of Agaricus blazei Murrill Mushrooms (mg 100g −1 dm)
The content of the PP vitamin (mg 100g −1 dm) in Agaricus blazei Murrill mushrooms ranged between 30.33 and 56 mg 100g −1 dm in 2017 and between 29 and 54.67 mg 100g −1 dm in 2018, depending on the experimental compost variants used. (Figure 7). In both experimental years the maximum value was recorded in the mushrooms harvested on the composite compost with the proteinaceous addition of corn flour (V9).   As with other B-complex vitamins, PP vitamin levels are higher in mushrooms grown on composite compost (C3), with or without protein addition. The highest value (55.33 mg 100 g −1 dm) was obtained with the use of the protein admixture of corn flour (A3).

Discussion
Compared with the Agaricus bisporus mushrooms, the ergosterol content of the Agaricus blazei Murrill mushroom is lower compared to the values presented by other researchers.
Vitamin C, is related to the C6 sugars, being the aldono-1,4-lactone of a hexonic acid (L-galactonic or L-gulonic acid) and contains an endiolic group at 2 and 3 carbon atoms. Its synthesis may be influenced by the presence of proteins [47][48][49], in the case of our study, the vitamin C content is influenced by the protein addition used in the culture substrate.
Vitamin C sometimes reacts with vitamin E, producing a vitamin C radical and regenerating vitamin E. Both radicals are poorly reactive species because of their unpaired electron [56,57].
Comparing the vitamin B1 content of Agaricus blazei Murrill mushrooms with other foods that are known thiamine sources, such as cereals, this is lower, but is close to those contained in vegetables and higher than in eggs. [58].
Regarding the vitamin B 1 content of Agaricus blazei Murrill mushroom, Tsai et al. [50] mention the amount of 720 µg 100 g −1 dm, Carneiro et al. [51] mention the amount of 940 µg 100 g −1 dm, and Cohen et al. [52] mention the amount of 1150 µg 100 g −1 dm. The values determined by us are found within these ranges.
Compared with the Agaricus bisporus mushrooms, the vitamin B1 content of the Agaricus blazei Murrill mushroom obtained by us is higher and is confirmed by other authors.
Regarding the B 2 vitamin content of the Agaricus blazei Murrill mushrooms, Tsai et al. [50] found the amount to be 3550 µg 100 g −1 dm, Carneiro et al. [51] found the amount to be 4625 µg 100 g −1 dm, and Cohen et al. [52] found the amount to be 6200 µg 100 g −1 dm. The values determined by us are found within these ranges.
Compared with the Agaricus bisporus mushrooms, the B 2 vitamin content of the Agaricus blazei Murrill mushroom has lower values. Thus, Mattila et al. [53] mention 5100 µg 100 g −1 dm, Furlani and Godoy [54] mention 2500 µg 100 g −1 dm, and Bernas and Jaworska [55] mention 1800 µg 100 g −1 dm Since the analyzed samples were from different growing substrates, different protein addition could have contributed to these differences. As mentioned by other authors, the different substrates used for cultivation could also have altered the composition of the mushrooms [59,60].
Regarding the vitamin B 9 content of Agaricus blazei Murrill mushroom, Cohen et al. [52] determined the value of 220 µg 100 g −1 dm, so the values determined by us exceed those found by other authors. In this case, the results obtained were influenced by the protein addition from the culture substrate. In the literature, we have not found data on the vitamin B 9 content in mushrooms grown on different substrates.
Detection of vitamin B 9 (Folic acid) in all experimental variants, within the range of 290 to 687 µg 100 g −1 dm in 2017 and from 277 to 657 µg 100 g −1 dm in 2018, shows that these mushrooms are rich in vitamin B9. Agaricus blazei Murrill mushrooms are a rich source of protein, vitamins, and minerals [61,62], showing a high interest as a source of raw material in the pharmaceutical industry, with folic acid being successfully used in the treatment of anemia and for pregnant women [63].
Analysis of vitamin B 12 in harvested mushrooms varied from variant to variant, with higher concentrations of B 12 detected in the outer peel than in the cap, stalk, or flesh, suggesting that vitamin B 12 is probably bacteria-derived [64]. Higher concentrations of vitamin B 12 were also detected in the mushrooms harvested from mixed substrate with protein addition (C3 A2, C3 A3). HPLC and mass spectrometry showed that vitamin B 12 retention time and mass spectra were identical to those of the standard B 12 vitamin and those of food products.
Regarding the vitamin B 12 content of the Agaricus blazei mushroom, they are found within the ranges found in the literature of Carneiro et al. [51], 920 µg 100 g −1 dm, and Cohen et al. [52], 760 µg 100 g −1 dm.
PP vitamin (B 3 -Niacin), contained in mushrooms, helps the human body to produce energy from blood sugar, helps to reduce the cholesterol levels, gives the skin a healthy and shiny texture, and stimulates the production of progesterone, estrogen, and testosterone. [65].
Regarding the PP vitamin content of the Agaricus blazei Murrill fungus, Tsai et al. [50] found the amount to be 45 mg 100 g −1 dm, Carneiro et al. [51] found the amount to be 53 mg 100 g −1 dm, and Cohen et al. [52] found the amount to be 36 mg 100 g −1 dm. The values we determined are found within these ranges.
Following the results presented above, it can be said that the consumption of mushrooms can increase the intake of vitamins in certain diets.
Following the evolution of the market for food supplements, especially those obtained from mushrooms with a scientifically proven therapeutic effect, it can be said that the demand for such food supplements is increasing. If it is desired to increase the content of certain vitamins in these food supplements, one can intervene on the substrate recipes, thus influencing the chemical composition and the content in bioactive substances of the finished product. In addition, if the products obtained from a specific recipe have a high level of bioactive substances, but also the level of production has a high biological efficiency, then that recipe can also be made at the industrial level, without causing significant losses, with the initial tests being carried out on small amounts of substrate.

Biological Material Used in Experiments
The biological material that has been used in the experimental trial comes from a pure culture and is represented by the M7700 strain. The species Agaricus blazei Murrill (Figure 8), is popularly called "the royal fungus", "God's mushroom", "the goddess's mushroom", and "the fungus of the sun".
The cap is thick, fleshy, hard, small to large in size, 5-11 cm in diameter, semi-globular at the beginning semi-globular then convex, smooth on the edges with pores in the center, white, yellow-creamy like an almond, or light brown to dark brown. On the edge of the hat are stuck pieces of veil. It has an almond flavor (Figure 9). The cap is thick, fleshy, hard, small to large in size, 5-11 cm in diameter, semi-globular at the beginning semi-globular then convex, smooth on the edges with pores in the center, white, yellowcreamy like an almond, or light brown to dark brown. On the edge of the hat are stuck pieces of veil. It has an almond flavor (Figure 9).    The cap is thick, fleshy, hard, small to large in size, 5-11 cm in diameter, semi-globular at the beginning semi-globular then convex, smooth on the edges with pores in the center, white, yellowcreamy like an almond, or light brown to dark brown. On the edge of the hat are stuck pieces of veil. It has an almond flavor (Figure 9).    The cap is thick, fleshy, hard, small to large in size, 5-11 cm in diameter, semi-globular at the beginning semi-globular then convex, smooth on the edges with pores in the center, white, yellowcreamy like an almond, or light brown to dark brown. On the edge of the hat are stuck pieces of veil. It has an almond flavor (Figure 9).   Basidiospores are dark brown, up to 6-8 × 4-5 microns, have a chocolate color, are wide-elliptical to oval, and have no epispore.
The stipe is short and thick, like a column, filled in, cylindrical, has a white color, and is attached to a mycelian base. On the touch by hand, the leg of the mushroom becomes yellow (Figure 11). A ring remains of the leg after the velvet breaks. The length is 6-13 cm and the diameter is 1-3 cm. The stipe is short and thick, like a column, filled in, cylindrical, has a white color, and is attached to a mycelian base. On the touch by hand, the leg of the mushroom becomes yellow (Figure 11). A ring remains of the leg after the velvet breaks. The length is 6-13 cm and the diameter is 1-3 cm.

Experimental Factors
To study the influence of compost recipes on the quality of mushroom production, as well as on the vitamin content of mushrooms, a bifactorial experimental trial was designed.
The combination of experimental factors resulted in 12 variants, presented in Table A1.
The compost recipes used in the experiment and the preparation process thereof are those described by Rozsa et al. [66]. (Table A2).
Considering that in the experiment factor A-the culture substrate, had 4 graduations, for the directed composting, a 4-composting box was designed to control, realize, and record the optimum environmental conditions for composting ( Figure A1, A2 a-e).
The stages of mushroom culture are presented in the appendix B, Figure B1-B6.
All reagents were provided by SC CIUPERCARIA SRL, Aghireșu-Fabrici, Cluj County Horticultural Products Laboratory, and they all were of reagent-grade purity.

Samples Preparation
Sampling was done for each experimental variant, formed from whole fruiting bodies of different sizes. These were washed with distilled water, cut into pieces, and frozen at −80 °C, then dried by lyophilization. The samples thus obtained were ground until a fine powder was obtained, which was kept in tightly sealed plastic bags at −20 °C until the analysis was performed.

Sample Extraction
For the extractions, the Pegg method [67] was used with some modifications. Thus, for each sample, 100 mL of ethanol and 10 g of powder were used in an Erlenmeyer flask covered with aluminum foil and stirred at 210 rpm for 80 min at 25 °C. After which, the extract was filtered and the residue was filtered. It was extracted two more times, then the extracts were mixed and the

Experimental Factors
To study the influence of compost recipes on the quality of mushroom production, as well as on the vitamin content of mushrooms, a bifactorial experimental trial was designed.
The combination of experimental factors resulted in 12 variants, presented in Table A1.
The compost recipes used in the experiment and the preparation process thereof are those described by Rozsa et al. [66] (Table A2).
Considering that in the experiment factor A-the culture substrate, had 4 graduations, for the directed composting, a 4-composting box was designed to control, realize, and record the optimum environmental conditions for composting ( Figure A1, Figure A2a-e).
The stages of mushroom culture are presented in the Appendix B, Figures A3-A8.
All reagents were provided by SC CIUPERCARIA SRL, Aghiresu-Fabrici, Cluj County Horticultural Products Laboratory, and they all were of reagent-grade purity.

Samples Preparation
Sampling was done for each experimental variant, formed from whole fruiting bodies of different sizes. These were washed with distilled water, cut into pieces, and frozen at −80 • C, then dried by lyophilization. The samples thus obtained were ground until a fine powder was obtained, which was kept in tightly sealed plastic bags at −20 • C until the analysis was performed.

Sample Extraction
For the extractions, the Pegg method [67] was used with some modifications. Thus, for each sample, 100 mL of ethanol and 10 g of powder were used in an Erlenmeyer flask covered with aluminum foil and stirred at 210 rpm for 80 min at 25 • C. After which, the extract was filtered and the residue was filtered. It was extracted two more times, then the extracts were mixed and the residual solvent was removed from the extracts in a rotary evaporator under reduced pressure at 40 • C until drying. The aqueous extract was lyophilized. The dried extracts obtained were used to make the determinations.

Methods Used for Analyses
The principles and procedures for selected vitamin analysis are described by Pegg and Eitenmiller [67].

Statistical Analysis
The processing of the obtained results was made by analyzing the polyfactorial variance, on each analyzed character, and the statistical interpretation was made with the STATISTICA 10 program with the Duncan test.

Conclusions
The nutritional value of the Agaricus blazei Murrill mushrooms, evaluated by their chemical composition and vitamin content, highlights the product obtained on the classic compost was improved with the protein addition of corn flour. Mushrooms harvested on this substrate have the highest levels of B1, B9, B12, PP, and vitamin Cs.
The content of ergosterol as a precursor of vitamin D2, which is generally higher in mushrooms than in other crops, has higher values in the product obtained on the classic compost, with the addition of corn flour and wheat barn.
Based on the results of the research and the general conclusions formulated, the following recommendations for production can be made: For Agaricus blazei Murrill mushrooms culture, synthetic wheat straw (66%), poultry manure (28%), gypsum (4%), and urea (2%) can be used with very good results. Depending on the local material resources in the area, it can be used as a culture substrate in the classical compost, consisting of garbage with bedding (93%), gypsum (5%), superphosphate (1%), and ammonium sulphate (1%).  Reed + horse manure (C4) corn flour 3% (A3)   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, built underground; 5heating elements for wetting water (purine); 6-water/purine recirculation pump; 7-recirculation Figure A1. Sketch for the composting facility-4 modules [68]. 1-tank with capacity of 1 m 3 for compost components; 2-rack for compost; 3-tank heating system for composting; 4-tank for the collection and recirculation the water excess, built underground; 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. 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.

Appendix B
The stages of mushroom culture

Appendix B
The stages of mushroom culture 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.

Appendix B
The stages of mushroom culture 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.

Appendix B
The stages of mushroom culture