**3. Material and methods**

The study was carried out at the Edible Mushroom Cultivation Laboratory from the Department of Forest Products in the "Instituto Nacional de Pesquisas da Amazônia" - INPA, in the following steps:

#### **3.1 Collection, drying and preparation of material**

Wood residue (sawdust) was chosen based on the generation of the wood waste produced by local lumber industry. Collection, drying and preparation of materials were done at CPPF/INPA, using sawdust of *Anacardium giganteum Hanck* ex Engl (cajuí). After the collection of the residues, they were dried (12% of humididy) in a solar dryer at CPPF/INPA, and packaged into plastic bags of 100 L until the preparation of the substrates.

#### **3.2 Production of a primary and secondary matrix and the "spawn"**

The strain of *Lentinus strigosus (Schwinitz) Fries* was taken from the collection of fungi at INPA Institute. Mycelial fragments of fungus (stored in test tubes) were transferred to a Petri dish containing malt medium and incubated at 27 ºC until colonization by the fungus (primary matrix) that was used as a source of inoculum for the secondary matrix . Mycelial disks, 9 mm in diameter, were removed from the primary matrix and transferred to Petri dishes containing SDA medium (sawdust-dextrose-agar), prepared according to Sales-Campos (2008), named secondary matrix. "Spawn" is the source for inoculation of the cultivation substrate, considered here as a tertiary matrix. This matrix was produced from cajuí sawdust, with humidification of 75%. The pH was corrected to approximately 6.5, by adding CaCO3. Then that substrate was deposited on glass bottles of 500 mL, in 200 g portions, which were autoclaved at 121 °C for 45 minutes. After cooling, the substrate was inoculated with the secondary matrix. The bottles were partially closed, and kept in special chamber with biochemical oxygen demand (BOD) at 25 ± 2 °C until the complete colonization of the substrate by fungus. This matrix served as a source of inoculation for the cultivation substrates for the production of *L. strigosus* mushrooms

#### **3.3 Preparation of cultivation substrate and processing**

The cultivation substrate was prepared from the same residue (cajuí sawdust) as the spawn inoculums. It consisted of 88% of sawdust + 10% of the soy bran as a protein source + 2% of CaCO3, for pH adjustment (6.5). The material was homogenised and humidified to 75%, and packed into bags of high density polyethylene-HDPE (1 kg capacity). Only 500 g of the substrate (wet basis) were put into each bag, with ten repetitions. The substrates were autoclaved at 121 °C for one hour. After that, they were cooled and inoculated with a tertiary matrix under axenical conditions. Each experimental unit (the bag containing the substrate) received 3% of the inoculum in relation to the wet weight of the substrate. They were taken to an incubating chamber until the colonization of the substrate by the fungus. Afterwards, they were transferred to a production chamber. The control samples were also prepared as above, but without inoculation by the fungus. The bags were taken to an oven with air circulation at 55 ± 5 ºC and dried to a constant weight, in order to obtain the dry mass of the initial substrate (DMIS) so that they were used to calculate the productivity, based on the biological efficiency index of substrate (BE) and the loss of organic matter (LOM).

#### **3.4 Experimental conditions**

244 Recent Trends for Enhancing the Diversity and Quality of Soybean Products

In Brazil, the necessary time for the complete development of the shiitake mushroom is not well defined due to climatic variations. The fruiting occurs over a period between three and twelve months after the inoculation, depending on the temperature of the region and the maintenance of moisture in the log (Eira and Minhoni, 1997; Eira and Montini, 1997). To accelerate this process in the cultivation on eucalyptus logs, the authors recommend soaking the miceliated logs for induction, after the incubation period when the first signs of primordia emission (callus or popcorns) which usually appear after 2 to 3 months. Mineral supplementation in water immersion increased the productivity of this mushroom. However, the increase of productivity and the efficiency of energy conversion were only possible in logs well colonized by the fungus (Queiroz, 2002; Eira and Minhoni, 1997; Eira

In relation to water temperature for immersion, there is a controversy, probably because of environmental differences, and observations often without experimental parameters (Eira and Minhoni, 1997; Eira and Montini, 1997). Some Brazilian producers who own cooling bath system report positive results since this system causes a steady temperature differential of 5-100 C. However, experiments performed in the Module of Mushrooms of the Faculty of Agricultural Sciences "Universidade Estadual Paulista" (FCA/UNESP), in Botucatu, São Paulo State, these same authors report that, in regions with mild climate and thermal amplitude greater than 100 C, the use of ice for cooling did not show significant difference in

Induction time depends on environmental conditions and age of the logs and the fruiting temperature varies from 5 to 300 C depending on the strain and the spawn used for cultivation. The relative humidity of the location of the logs should be between 80 and 90%. The emergence of primordia will be within two to three days and harvesting can be made after seven to ten days, and in cool seasons the metabolism of the fungus is reduced, increasing the time before harvest (Eira and Minhoni, 1997). The induction bath can be done in stages, depending on the needs of the producer, thus inducing bath of logs can be programmed as a function of demand (Eira and Minhoni, 1997; Eira and Montini, 1997).

The study was carried out at the Edible Mushroom Cultivation Laboratory from the Department of Forest Products in the "Instituto Nacional de Pesquisas da Amazônia" -

Wood residue (sawdust) was chosen based on the generation of the wood waste produced by local lumber industry. Collection, drying and preparation of materials were done at CPPF/INPA, using sawdust of *Anacardium giganteum Hanck* ex Engl (cajuí). After the collection of the residues, they were dried (12% of humididy) in a solar dryer at CPPF/INPA, and packaged into plastic bags of 100 L until the preparation of the substrates.

The strain of *Lentinus strigosus (Schwinitz) Fries* was taken from the collection of fungi at INPA Institute. Mycelial fragments of fungus (stored in test tubes) were transferred to a Petri dish containing malt medium and incubated at 27 ºC until colonization by the fungus

and Montini, 1997).

relation to normal bath immersion.

**3. Material and methods** 

INPA, in the following steps:

**3.1 Collection, drying and preparation of material** 

**3.2 Production of a primary and secondary matrix and the "spawn"** 

The experiment was conducted indoors. The bags contained the substrates were incubated in a climatic chamber at the temperature of 25 ± 3 °C, in the absence of light and at around 80-85% humidity, in order to allow substrate colonization until the production of primordia. Then, they were transferred to the production chamber. The temperature was reduced from 25 ºC to 22 ºC to induce primordial emission and to allow the production of basidioma (fruit body of the mushrooms) in a way that it would be as uniform as possible. Light intensity was maintained at 2000 Lux, with a photoperiod of 12 hours per day. The relative humidity was scheduled to 95% during the "fructification. The total period of cultivation was 100 days. After "fructification", the mature mushrooms were collected and weighed, and then oven dried for the determination of moisture, dry mass and chemical analyzes. During cultivation, the variables analyzed were: biological efficiency (BE), and loss of organic matter (LOM). Biological efficiency (used to express the productivity of fungus), was calculated according to Tisdale et al. (2006) and Das and Mukherjee (2007):

$$BE = \frac{FMM}{DMS} \ast 100$$

Productivity and Nutritional Composition of *Lentinus strigosus* (Schwinitz)

reported in this study (Table 3).

Substrate Cajui Sawdust

**5. Conclusions** 

protein for the fungus.

**6. References** 

168050-9, New York.

Fries Mushroom from the Amazon Region Cultivated in Sawdust Supplemented with Soy Bran 247

Özçelik and Pekşen (2007), analyzing the application of hazelnut shells in the formulation of substrate for mushroom cultivation *Lentinula edodes*, reported that the biological efficiency of the substrate made with hazelnut shells only, was considered to be low (43.73%). However, when the proportion of hazelnut shells was reduced and combined with wheat straw (25:75) the biological efficiency was considered good (62.24%). The result however, is less than that

Philippoussis et al. (2007) tested the productivity of agricultural residues (sawdust of oak, wheat straw and corncobs) in the cultivation of *Lentinula edodes* and found that corncobs and wheat straw presented higher rates of biological efficiency: 80.64% and 75.23% respectively, which were similar to those presented in this research. Alberto and Lechner (2007) however,

Royse and Sanchez (2007) tested three formulations for the cultivation of *L. edodes*, combining wheat straw and oak residues. They found that the substrate with higher proportions of wheat straw (in relation to oak residue), provided the best biological efficiency (98.9%) at the end of 4 harvests. These results are superior to the ones obtained in this work. However, 80% of Biological Efficiency presented by *L. strigosus* cultivated in cajuí

Productivity Results Nutritional Composition

Total Protein (%)

Total Fiber (%)

Lipid

Ash

(%)

(%)

Loss of Organic Matter (LOM) (%)

Table 3. Results of Productivity and Nutritional Composition of the edible mushroom *Lentinus strigosus* cultivated on cajuí wood waste, supplemented with soy bran.

Standard deviation 6.94 4.85 1.00 2,00 0.30 1.00 Average **80.0 47.51 20 18 2.5 5** 

The high biological efficiency of the mushroom in this substrate, formulated with the cajuí sawdust supplemented with soy bran, makes its use feasible for the cultivation of *Lentinus strigosus* mushroom from the Amazon Region. The soy bran provided a good source of

The findings presented herein point out the utilization of the Amazon wood waste as substrate for the mushroom cultivation, which will certainly promote the improvement of the social and economical conditions of its people and the sustainability of the biodiversity

*L. strigosus* can be considered an important food in terms of their characteristics: rich in

Bononi, V. L.; Capelari, M.; Maziero, R. & Trufem, S. F. B. S. (1999). *Cultivo de cogumelos* 

Crisan, E. V.; Sands, A. (1978). A nutritional value, In: *The biology and cultivation of edible* 

*mushroom*, Chang, S. T.; Hayes, W.A. (Eds), pp.137-168, Academic Press, ISBN: 0-12-

resources, enabling the establishment of a new economical niche in the region.

*comestíveis*. 2nd Ed., Ícone , ISBN: 85-274-0339-0, S. Paulo.

protein and low in fat, important for nutrition and human health.

obtained lower BE (61.93%), cultivating *Lentinus tigrinus* with Salix sawdust.

sawdust supplemented with soy bran in the present study is considered high.

Biological Efficiency (BE) (%)

Where: BE= Biological efficiency, % FMM= Fresh mass of mushrooms, g DMIS= Dry mass of the initial substrate, g

The loss of organic matter (LOM) is the index that evaluates the substrate decomposition by the fungus. It was evaluated according to Sturion (1994), expressed by the following formula:

$$LOM = \frac{DMIS - DMSSS}{DMSS} \ast 100$$

Where:

LOM = loss of organic matter,% DMSS = Dry mass of the spent substrate, g DMIS = Dry mass of the initial substrate, g
