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Physiologic response of Agaricus subrufescens using

different casing materials and practices applied inthe cultivation of Agaricus bisporus

Eust�aquio Souza DIASa, Diego Cunha ZIEDa,b,*, Danny Lee RINKERc

aUniversidade Federal de Lavras, Departamento de Biologia, CP 3037, 37200-000 Lavras, MG, BrazilbFaculdades Integradas de Bauru (FIB), Rua Rodolfina Dias Domingues, 11, Jardim Ferraz,

17056-100 Bauru, SP, BrailcUniversity of Guelph e Vineland Campus, 4890 Victoria Avenue North, P.O. Box 7000, VinelandStation,

Ontario, Canada L0R 2E0

a r t i c l e i n f o

Article history:

Received 11 April 2013

Received in revised form

19 June 2013

Accepted 21 June 2013

Available online 1 July 2013

Corresponding Editor:

Marc Stadler

Keywords:

Agaricus blazei

Agaricus brasiliensis

CACing

Re-casing

Ruffling

* Corresponding author. Coordenac~ao AgronoSP, Brazil. Tel.: þ55 14 2109 6200; fax: þ55 14

E-mail address: dczied@gmail.com (D. C.1878-6146/$ e see front matter ª 2013 The Bhttp://dx.doi.org/10.1016/j.funbio.2013.06.007

a b s t r a c t

Casing materials and practices used in the cultivation of Agaricus bisporus were evaluated

in the cultivation of Agaricus subrufescens, using the best techniques for optimization of

production, including the possibility of re-casing of the compost for the production of a sec-

ond crop of mushroom. Casing based on peat moss, loam soil or coir was compared to cas-

ing material mixed with or without spawn-run compost. Based on the results, we conclude

that the casing layer used in the cultivation of A. subrufescens should not necessarily be the

same as that used in the cultivation of A. bisporus. For the tested strain cultivated with

loam soil as casing layer, the ruffling technique is highly superior to CACing and should

be pursued in further research. The re-casing of compost in new cycles showed good re-

sults suggesting that the currently used compost could be improved.

ª 2013 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Introduction Brazil (Uryu 1995; Braga et al. 1998) with Eira (Eira 2003; Eira

Agaricus subrufescens Peck, also known as Agaricus brasiliensis

Wasser et al. and Agaricus blazei (Murrill) ss. Heinemann

(Wasser et al. 2005; Kerrigan 2005; Eira et al. 2005a; Arrillaga

& Parra 2006; Dias et al. 2008; Cappelli 2011; Maia et al. 2012)

has aroused great interest in various parts of the world

because of its medicinal properties, which is used in the pre-

vention or treatment of diabetes, atherosclerosis, hepatitis,

hypercholesterolaemia, and heart disease, among others

(Zied 2011). Cultivation of A. subrufescens was first studied in

mia (FIB/BAURU), Rua Ro2109 6202.Zied).ritish Mycological Societ

et al. 2005b; Dias 2010) pioneering the research on the cultiva-

tion of this mushroom in Brazil.

Many technologies used in the cultivation ofA. subrufescens

are adopted from the production of Agaricus bisporus. The cas-

ing layer is one of these steps and is considered by some

authors as ‘the skin of the substrate’ (Vernooij 2008), one of

the major influences on yield and final quality of these mush-

rooms (Pardo et al. 2003).

Various casingmaterials are used in A. bisporus cultivation.

Casing materials are mixtures of water holding materials and

dolfina Dias Domingues, 11 e Jardim Ferraz, CEP 17056-100, Bauru,

y. Published by Elsevier Ltd. All rights reserved.

570 E. S. Dias et al.

lime for pH adjustment. The most common starting material

is peatmoss inmanymushroomgrowing regions of theworld.

Loam soil, weathered and/or composted spent Agaricus com-

post (Rinker 1993), and coconut coir (Noble 2011) have been

used successfully.

Casing management techniques have emerged that have

resulted in increased yield, better management of the unifor-

mity of the Sporocarp flush and individual sporocarp size and

shorten time to the 1st flush and length of total crop. These

procedures include scratching, ruffling, and adding compost

colonized by the mycelium to the casing (a.k.a. CACing; com-

post added at casing).

The casing surface is ‘scratched’ once the initial build-up of

casing water has been accomplished and the mycelium has

begun to grow up into the casing. When the casing is

scratched, care is taken not to disturb the mycelia. The func-

tion of scratching is to break up the surface compaction, re-

duce the CO2 level and redistribute the casing material over

thinner casing areas (MacCanna 1986).

‘Ruffling’ disturbs the entire casing layer and themycelium

that has grown up into it. Commercially, a specially designed

rototiller-like machine mixes the casing and the mycelium

when the mycelium has developed approximately three-

fourths into the casing layer (MacCanna 1986). Level compost

is required; otherwise, compost is mixed with casing in spots,

creating dense mycelial growth and reduced production in

those areas.

Two procedures addmycelium directly to the casingmate-

rial at casing time. The ‘CACing’ technique involves the mix-

ture of small amounts of compost fully colonized (spawn-

run compost) by the mushroom mycelium into the casing

layer at casing time (Zied et al. 2010a). Or, a commercially pre-

pared ‘casing inoculum’, a spawn-like material, is mixed with

the casing prior to its placement on the compost.

The present study evaluated the practices of CACing and

ruffling, as it is used in the cultivation of A. bisporus, on the

cultivation of A. subrufescens using the best techniques for op-

timization of production, including the possibility of reuse of

compost on new crops.

Materials and methods

Cultivation process

The experiments were conducted at the Vineland Mushroom

Research Facility on the Vineland Campus of University of

Guelph, Vineland, Ontario, Canada. Commercially prepared

Agaricus bisporus Phase I compost was provided by Greenwood

Mushroom Farm, Ashburn, ON. The development of compost

occurred in the following stages: 7 d of pre-wetting straw

(bundles maintained), 4 d of Phase I in bunker with forced

air flow, 4 d in another bunker, and 4 d in a final bunker.

The compost was pasteurized (8 h at 60 �C) and conditioned

(4 d at 45e50 �C) in 5 d using amini-phase II tunnel at Vineland

(Rinker 1993). Compost (26 kg; fresh wt) was inoculated with

Agaricus subrufescens CS1 (Brazilian commercial strain, iso-

lated from growers of the City of Piedade, deposited in Univer-

sidade Federal de Lavras, Department de Biology, CP 3037,

37200-000, Lavras (MG), Brazil) at the rate of 1.3 % (wt:wt)

wheat spawn and placed into 0.25 m2 plastic trays (Zied et al.

2012). Then, the trays were incubated for 15 d at 25 �C, relativehumidity of 95 % and in darkness. After the incubation period

(spawn-run), the compost surface was top dressed with peat

moss, loam soil or coconut coir.

Preparation of the casing materials

In a sanitized environment, 54 kg of commercial peat moss

were mixed with 10 kg of agricultural lime, placed into perfo-

rated bins (46 � 46 � 26 cm) and steamed pasteurized for 12 h

at 60 �C. The pH value at casing was 7.85. Local loam soil was

air-dried and sieved to avoid diameter particles greater than

10 mm and, after that, 311 kg of soil were mixed with 5.2 kg

of agricultural lime and 120 L of water. The mixture was

placed into perforated bins (46� 46� 26 cm) and steamed pas-

teurized for 12 h at 60 �C. The pH value at casingwas 8.0. Bricks

of coconut coir (total ¼ 28.3 kg) were submerged into water

(60 L) overnight. The bricks absorbed all the water and it was

necessary to add an additional 180 L of water. Thewet coconut

coir (267 kg) was then mixed with 10 kg of agricultural lime,

placed into perforated bins (46� 46� 26 cm) and steamed pas-

teurized for 12 h at 60 �C. The pH value at casing was 8.2.

Experimental design

The 1st and 2nd experiments were two-way factorial designs

(three casingmaterials and two techniques) with five replicates

per treatment (a tray of 26 kg of fresh wt of compost). Ten trays

were cased with peat moss (5.5 kg/tray), loam soil (15.5 kg/tray)

or coconut coir (6.9 kg/tray) in order to have a 5 cm casing layer.

In the 1st experiment, half of each set of trays were CACed

by mixing 300 g of manually shredded spawn-run compost in

the casing of each tray. In the 2nd experiment, half of each set

of trays were ruffled 6 d after casing by manually mixing the

casing material in situ down to the compost surface. For both

experiments, the casing of the other half of the corresponding

trays were not disturbed and served as a control.

After these two experiments, the 3rd experiment were

done, with a completely randomized design with five repli-

cates per treatment (a tray of fresh wt of 26 kg of compost)

where peat moss were added (re-casing) over the old compost

(casings used in 1st and 2nd experiment were removed for the

production of a second crop of mushroom) and subjected to

pinning in two temperatures (19 and 25 �C); as a control,

a new compost were used for pinning at 19 �C with two con-

centration of peat moss and lime (1:1 and 5:1, vv) (six

treatments � five replicates ¼ 30 experimental units). Also

Ruffing technique was used in 3rd experiment (Table 1).

Cultural conditions

Cased trays were randomized and arranged in two tiers, three

trays high with a maximum of six trays per level per tier.

During the pre-production period (case-run), the compost tem-

perature was maintained at 25 �C with a relative humidity of

95 % and a carbon dioxide level of 3000 ppm. Primordia were

stimulated by decreasing the air temperature to 19 �C and the

carbon dioxide level to about 1000 ppm. During production,

the air temperature was maintained at 25 �C with relative

Table 1 e Materials used as casing layer, aim and other relevant information used in each experiment.

Experiment Casing Aim Harvest time Note

1 Peat moss, soil and

coconut coir

Check the effect of CACing technique 120 d 300 g of spawn-run compost were

used per trays

2 Peat moss, soil and

coconut coir

Check the effect of ruffing technique 120 d The ruffing was performed 6 d

after casing

3 Peat moss Check the effect of temperature in

the pinning, the amount of lime added

on casing and the re-casing

of compost for the production of a

second crop of mushroom

84 d Pinning at 25 �C and 19 �CPeat:Lime (1:1 and 5:1, v:v)

A. subrufescens cultivation with A. bisporus tech. 571

humidity of 95% and carbon dioxide level of 1000 ppm. The cul-

tural environment was controlled by a computerized system.

Casing was irrigated as required to maintain maximum water

holding capacity.

Evaluation of data

Sporocarps were harvested before the cap opening, counted

and weighed daily. Yield was calculated as fresh weight of

sporocarp divided by fresh weigh of compost multiplied by

100, and expressed as %. The earliness was calculated as the

number of days between casing and the 1st harvest (Mamiro

& Royse 2008). Precociousness (yield during first half of har-

vest interval divided by total yield multiplied by 100,

expressed as %) was calculated (Zied et al. 2010b).

Statistical analysis

Data were analyzed using the Proc Glm Procedure of SAS (SAS,

Inc., Cary, NC) with means separated by Tukey test (P � 0.05).

Relationships between variables were determined by Pearson

product-moment correlation analysis (SAS, Inc, Cary, NC).

Results

1st experiment e addition of spawn-run compost to casing

The type of casingmaterial significantly affected productivity,

number of sporocarps, individual sporocarp weight,

Table 2 e Effect of CACing technique on agronomic behaviour

Casing Yield, % Number mush., unit We

CACing Without CACing Without CAC

Peat 6.8a A 7.2a A 48.5a A 52.1a A 38.9a

Soil 6.1ab A 8.0a A 44.8ab A 68.0a A 36.3a

Coir 2.1b A 1.8b A 11.5b A 11.1b A 45.9a

CV 58.33 69.38 69.38

LSD 4.47a 3.70b 36.64 30.34 36.64

Media 5.3 39.36 39.36

Lowercase letters compare results in the same columns within each varia

same line within each variable analyzed.

CACing ¼ compost added to casing; CV ¼ coefficient of variation.a LSD ¼ low significance difference of CACing values.b LSD ¼ low significance difference of casing values.

precociousness, and earliness (Table 2). Coconut coir was an

inferior casing material as it related to productivity, number

of sporocarps, precociousness, and earliness. Soil and peat

moss were statistically equal in yield, number of sporocarps,

individual sporocarp weight, and earliness. Peat moss and co-

conut coir were superior to soil in precociousness with nearly

20 % more of the total sporocarp yield harvested during the

first half of the harvest period.

CACing significantly affected positively the precociousness

and negatively the earliness. Thirteen percent more yield was

harvested using the CACing technique during the first half of

harvest. However, time to first harvest was significantly

delayed in the CACing technique by 7 d.

The only significant interaction (0.0436) was with the vari-

able earliness (Table 5). CACing more seriously delayed onset

of first harvest when coir was used as a casing material than

with either soil or peat moss (Table 5).

Yield was positively correlated with number of sporocarps

and negatively correlated with individual sporocarp weight.

More sporocarps produced higher sporocarp yield but with in-

creased yield individual sporocarpweight decreased. Earliness

was negatively correlatedwith increased productivity, i.e., the

sooner sporocarps began to be harvested the greater the total

yield.

2nd experiment e ruffling the casing layer

The type of casingmaterial significantly affected productivity,

number of sporocarps, individual sporocarp weight,

of A. subrufescens (1st experiment).

ight mush, g Precociousness Earliness

ing Without CACing Without CACing Without

A 52.1a A 84.2 b A 61.9a B 30.3a A 26.4a A

A 68.0a A 57.2a A 52.5a A 27.5a A 24.3a A

A 11.1b A 81.7b A 69.9a A 39.6b B 25.4a A

23.17 18.72

30.34 22.42 18.56 7.74 6.41

67.95 29.04

ble and treatment analyzed. Uppercase letters compare results in the

Table 3 e Effect of ruffling technique on agronomic behaviour of A. subrufescens (2nd experiment).

Casing Yield. % Number mush. unit Weight mush. g Precociousness Earliness

Ruffling Without Ruffling Without Ruffling Without Ruffling Without Ruffling Without

Peat 5.8b A 4.1b A 45.0b A 26.5b A 36.0b A 45.5a A 59.7a A 61.5a A 34.0a A 38.9b B

Soil 18.7a A 9.8a B 164.0a A 74.8a B 30.6b A 35.5a A 60.6a A 68.2a A 30.7a B 22.1a A

Coir 1.3c A 1.3b A 5.6b A 6.6b A 68.6a A 51.9a B 88.5a A 85.3a A 35.1a B 26.7a A

CV 41.85 54.56 25.98 27.56 11.75

LSD 4.10a 3.39b 41.78 34.59 16.55 13.70 27.74 22.97 5.23 4.33

Media 6.8 53.77 44.74 70.69 31.29

Lowercase letters compare results in the same columns within each variable and treatment analyzed. Uppercase letters compare results in the

same line within each variable analyzed.

CV ¼ coefficient of variation.a LSD ¼ least significant difference of casing values.b LSD ¼ least significance difference of Ruffling values.

572 E. S. Dias et al.

precociousness, and earliness as in experiment 1 (Table 3).

Soil was statistically superior to peat moss and coir in produc-

tivity, number of sporocarps and earliness and equal to peat

moss in individual sporocarp weight, and precociousness. Al-

though coconut coir was superior to soil and peat moss in in-

dividual sporocarp weight and precociousness, its low yield

deems it an inferior casing material.

Ruffling the casing soil significantly increased yield by 40 %

and nearly doubled the number of sporocarps. Neither indi-

vidual sporocarp weight nor precociousness was significantly

affected by this technique. However, time to first harvest (ear-

liness) was delayed by 8 d when the casing was ruffled.

There were significant interactions between the type

of casing and whether or not the casing was ruffled for pro-

ductivity, number of sporocarps, individual sporocarp weight,

and earliness. Ruffling with soil casing nearly doubled yield in

comparison to the minor negative or positive effect in the

other two casing types.

The number of sporocarpswas nearly 120 % greater for ruf-

fled soil and 70 % for ruffled peat moss compared to only

a slight decrease in numbers for ruffled coconut coir. Individ-

ual sporocarp weight increased for ruffled coconut coir but

Table 4 e Effect of temperature of pinning and amount of lime o

Treatment Yield, % Number ofsporocarp, u

25 �C ruffling 15.35a 172.0ab

25 �C without ruffling 15.22a 188.3ab

19 �C ruffling 17.00a 195.8ab

19 �C without ruffling 14.27a 317.5a

New compost regular limea 3.92b 22.3b

New compost low limeb 6.67b 46.0b

CV, % 17.48 72.7

LSD 3.70 200.5

Media 12.07 157.0

Values followed by a different letter are significantly different at 5 % leve

LSD: least significance difference.

CV: coefficient of variation.a Peat:Lime (1:1, v:v).b Peat:Lime (5:1, v:v).

decreased for peat moss and soil. Time to first harvest (earli-

ness) was delayed by ruffling for coconut coir and soil casing

types, but shorten for peat moss casing.

Yield was positively correlated with number of sporocarps

and yield was spread more over the entire harvest interval

(precociousness), sic, a negative correlation. However, with in-

creased yield and number of sporocarps, individual sporocarp

weight was significantly lower (Table 5).

3rd experiment e use of old compost

The temperature of pining influenced the number of sporo-

carp, precociousness, and earliness and the amount of

lime influenced the precociousness and earliness (Table 4).

Regarding the influence of temperature of pining in the agro-

nomic behaviour of Agaricus subrufescens kept constantly at

25 �C reduces the time for first flush was observed; on the

other hand when the temperature was reduced to 19 �C for

the pinning a high precociousness was verified.

Table 5 explains the correlation between high yield with

lower weight of sporocarp (r ¼ �0.803) and the lower weight

n agronomic behaviour of A. subrubescens (3rd experiment).

Weight ofsporocarp, g

Precociousness, % Earliness,days

23.3b 43.4a 22.6a

23.0b 39.0ab 22.9a

22.9b 53.5a 24.0ab

16.6b 53.8a 24.4 ab

47.4a 33.9ab 34.5c

39.9 a 14.1b 32.5bc

20.8 39.6 19.3

10.5 27.6 19.3

28.9 39.6 26.8

l according to Tukey’s HSD test.

Table 5 e Correlations between the agronomic behaviour of Agaricus subrufescens.

Person correlation Number of sporocarp Weight of sporocarp Precociousness Earliness

1st experiment (CACing)

Yield 0.984 �0.393 �0.184 �0.440

<0.001 0.017 0.283 0.007

Number of sporocarp e �0.465 �0.234 �0.449

0.004 0.170 0.005

Weight of sporocarp e e 0.182 �0.081

0.287 0.636

Precociousness e e e 0.311

0.064

2nd experiment (ruffing)

Yield 0.984 L0.605 �0.435 �0.210

<0.001 <0.001 0.008 0.218

Number of sporocarp e L0.609 �0.385 �0.191

<0.001 0.020 0.264

Weight of sporocarp e e 0.264 0.314

0.119 0.062

Precociousness e e e �0.062

0.718

3rd experiment (temperature of pinning and amount of lime)

Yield 0.531 L0.803 0.419 L0.701

<0.001 <0.001 0.010 <0.001

Number of sporocarp e L0.794 0.482 �0.394

<0.001 0.002 0.017

Weight of sporocarp e e �0.473 0.628

0.003 <0.001

Precociousness e e e �0.470

0.003

First data correspond to r values (degree of significance) and second data correspond to the P values (probability). Values in bold represent a strong

correlation (r ¼ 0.6 to 0.8 and P < 0.001) and very strong correlation (r > 0.8 and P < 0.001).

A. subrufescens cultivation with A. bisporus tech. 573

of sporocarp with large number of sporocarp harvested

(r ¼ �0.794) obtained.

Comparing the results obtained in the 3rd experiment with

the 2nd experiment using the same casing (peat moss e with

andwithout ruffling), the yieldwas higher and precociousness

and the time for first flush (earliness) were lower in 3rd exper-

iment. Other interesting point is the fact that the high amount

of lime increases precociousness and reduces the time for first

flush, which probably occurred due to changes in the values

related to the cation exchange capacity (CEC).

Definitely re-casing old compost (‘spent’ mushroom com-

post) for the production of a second crop of mushrooms had

good results with yield, high number of sporocarps harvested,

reduced time for first flush and regular precocity (about�55%)

when pining at 19 �C were used.

Discussion

This study evaluated materials and practices used in the cul-

tivation of Agaricus bisporus for the cultivation of Agaricus sub-

rufescens. We selected casing materials based on peat moss,

coconut coir, or soil which is used commonly in Brazil. Ruf-

fling and CACing are successful common practices in the cul-

tivation of A. bisporus and were evaluated in conjunction with

alternative casing materials.

In both experiments, undisturbed soil ranked generally

higher in yield than peat moss or coir, in agreement with

the normal use of soil as a casing in Brazil. Soil was used

commonly in the A. bisporus sporocarp industry until the

1970s when peat moss began to gradually replace it. In the

A. bisporus industry the casing formulation now has evolved

away from use of 100 % peat moss to a blend of peat and other

materials. Increased density of the casing through additions of

sugar beet lime, for example, has maintained the water hold-

ing capacity, gas exchange and the pest free qualities of peat

while increasing the density of the casing facilitating better

quality sporocarps. A dense casing appears to be desirable

for A. subrufrescens. Research into blends of soil or other prod-

ucts with the benefits of peat should be explored.

The CACing technique showed no overall advantage in

productivity. However, this technique directly influenced the

precociousness values. The sooner more sporocarps are har-

vested is very critical considering that the commercial cultiva-

tion of A. subrufescens has a long harvest period under high

production temperatures (25e30 �C) and high relative humid-

ity (85e95 %), which results in high levels of diseases and in-

sects. In the cultivation of A. bisporus harvest time has

decreased to two weeks. With this shorten time, insect and

disease pests are substantially reduced or non-existent. So,

a technique that produces more sporocarps sooner is desir-

able from a pest management perspective (Rinker 1993).

And, these gains are most evident for the casings based on or-

ganic material rather than mineral. Zied et al. (2009) studied

different casings based on mineral soil and found precocious-

ness to increase from 51 % to 75 % based on addition of 0.5 %

acrylamide gel. However, in this study the advantage of preco-

ciousness is confounded by significant delay in first harvest

574 E. S. Dias et al.

(earliness). A delay to harvest increases the time for reproduc-

tion of insects andmites, as well as increases the grazing time

for damaging fly larvae. In contrast, the addition of spawn-run

compost or commercially prepared casing inoculum in the A.

bisporus industry has shortened the time to first harvest by

about 7 d.

Redistributing themycelia throughout the casing using the

ruffling technique was advantageous in yield and number of

sporocarp while slightly delaying first harvest. The effect of

ruffling soil was remarkable with twice the productivity, how-

ever, with 14 % lower individual sporocarp weight (2nd exper-

iment). The increased number of sporocarps (ca 120 %) will

make for individually lighter sporocarp weight. In A. bisporus

production ruffling helps produce uniform sporocarp size

and enables the entire flush of sporocarps to be harvested in

a shorter time. The uniformity of the flush can negatively af-

fect the individual sporocarp weight. Sporocarp weight is an

extremely important factor in grading for sale, i.e., sporocarps

with high weight are more valued. And, if the casing is ruffled

when there is very little mycelial growth or toomuchmycelial

growth, timing to first harvest will be delayed. With soil hav-

ing responded positively to ruffling, research into the influ-

ence of ruffling on individual sporocarp-by-sporocarp size

and the effect on the timing of the ruffling need to be further

explored.

Sporocarp yield (ranging in undisturbed soil from ca

8e10 %) was lower in these experiments than those values

(10e20 % over 120-d harvest period) obtained by Brazil’s

growers and by research conducted by Siqueira et al. (2009),

Colauto et al. (2010) and Zied et al. (2010b). Materials and

strains were different between our research and those exper-

iments conducted in Brazil. Zied et al. (2011) evaluated differ-

ent strains finding substantial differences in production on

the same substrate, e.g., French strain CA 487 (18.6 % produc-

tivity), hybrid CA 454 � 487 (11.2 %), Brazilian commercial

strain 99/30 (3.2 %) and a wild Brazilian strain CA 454 (2.3 %).

Llarena Hern�andez et al. (2011) also achieved high yield using

the French strain (CA 487) in cultivation with 65 d of harvest

phase.

At the end of these studies (1st and 2nd experiment) the

casing layer were removed and the old compost were re-

casing for the production of a second crop of mushrooms to

better understanding the low yield obtained and the possible

process of biodegradation of the compost. So we conducted

a third experiment using the same compost (old compost)

and trays used in 1st and 2nd experiment.

The reducing the temperature to the pinning does not af-

fect the agronomic performance of A. subrufescens except the

earliness values. Regarding the use of new compost, stay the

question about the use of A. bisporus compost in the cultiva-

tion of A. subrufescens.

It’s amazing emphasize the high yield achieved with the

old compost, which demonstrates the lack of cultivation tech-

nology for the production of this sporocarp, which we sup-

posed to think that will be the formulation of the compost

incorrect? The strain used in this experiment (related to trop-

ical climate) is not adequately for specific conditions tested? It

would be important to do a process of adaptation between

strains and compost? or What biodegradation processes

were influenced in old compound, the amount of lignin,

cellulose, hemicellulose, humic acid, soluble sugars, the C:

N: P ratio, and others chemical characteristics?

Conclusions

Based on these studies we conclude that the casing layer

used in the cultivation of Agaricus bisporus should not neces-

sarily be the same as that used in the cultivation of Agaricus

subrufescens. Casing based on soil showed the best yield in

comparison to peat moss or coconut coir. The ruffling tech-

nique is highly superior to CACing and should be pursued in

further research. The re-casing of compost in new cycles

showed good results suggesting that the currently used com-

post could be improved.

Acknowledgements

We would like to thank the Foundation for Research Support

of the State of Minas Gerais (FAPEMIG e CAG/BPD 00081-11)

and Foundation for Research Support of the State of S~ao Paulo

(FAPESP e 2012/15101-4) for financial support.

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