Cultivation of oyster mushroom (Pleurotus florida) on date palm residues in an environmentally controlled conditions in Saudi Arabia.
The utilization of agricultural wastes in a sustainable and an environmentally controlled manner has brought great attention in many parts of the world. Saudi Arabia is one of the leading date palm producing countries in the world where the number of palm trees is estimated around 23 million and this number is increasing gradually . Date palm produces large quantity of horticultural wastes. For instance, each tree produce about 10-20 leaves yearly . Other wastes like date pits represent an average of 10% of the date fruits. In general, the total quantity of date palm wastes in the Kingdom in 2008 was estimated around 656, 832 tonnes with an average of 2.89 tonnes/hectare . These useless date palm wastes are sources of many hazards like fire, host for many insect-pests and diseases . Although the palm wastes comprise cellulose, hemicelluloses, lignin and other compounds, which could be used in many biological processes, they were generally used as burning wood or burned in farms causing severe danger to the local environment [38,35,6].
The bioconversion of agricultural residues into food has attracted the world attention in recent years . The cultivation of edible mushrooms has a great potential for the production of protein rich quality food and for recycling of cellulosic agro- residues and other wastes [45,10,17] and serves as the most efficient and economically viable biotechnology for the conversion of lignocellulosic waste materials to high quality food [21,22]. They can easily grow on almost all types of cellulosic residues such as banana leaves, dried paddy straws , cotton waste and rice straws , sawdust enriched with poultry droppings , Jatropha  and even invasive weed species .
Mushrooms are fungi that produce lignocellulosic enzymes, which degrade complex organic matter and absorb the soluble substances [16,13,40]. Oyster mushrooms are one of the most popular edible mushrooms and belong to the genus Pleurotus and the family Pleurotaceae . The genus Pleurotus, is widely cultivated due to its favorable organoleptic and medicinal properties, simple and low cost production technology and higher biological efficiency . Oyster mushroom (Pleurotus spp.) is in the second position (after white button) in the world and its cultivation has increased rapidly during the last decade [42,44]. Oyster mushroom cultivation can play an important role in managing organic residues whose disposal has become a problem [18,28,49]. Furthermore, mushroom spent (grown media with mycelium) can be used as an organic fertilizer  or animal/fish feed [39,38].
Pleurotus florida is an edible mushroom having excellent flavor and taste  and is increasingly becoming popular as protein-rich delicious vegetable [33,41]. Pleurotus florida cultivation is popular due to low cost technology and easy availability of various substrates for its cultivation ([12,25,40]. It is widespread in temperate, subtropical and tropical zones and it shows the highest yield among the Pleurotus species . Therefore, the objective of this study was to evaluate the cultivation of oyster mushroom (Pleurotus florida) on date palm wastes in a controlled environment in Saudi Arabia.
MATERIALS AND METHODS
The study was conducted in the laboratories of Agricultural Engineering Department, College of Food and Agriculture Sciences, King Saud University, Riyadh City, Saudi Arabia, from August to November 2014.
Stock Pure Culture:
The pure culture of Pleurotus florida maintained on potato dextrose agar (PDA) was obtained from Dr. Renato G. Reyes, Center for Tropical Mushroom Research and Development, Central Luzon State University, Science City of Munoz, Nueva Ecija, Philippines. The culture was maintained at 4[degrees]C and recultured bimonthly.
Preparation of Spawn:
Wheat grains were boiled until tender (10-15 min) at a ratio of 1:1 (wheat grains: water) and allowed to cool down for 15 min, then the water was drained and the boiled grains were dried on cotton cloth. Then, the grains were packed tightly into the polypropylene bags (30 x 14 cm). One bag contained 350g of the prepared grains. The opening of the bags were plugged with cotton and covered with aluminum foil. The packed bags were sterilized in an autoclave at 121[degrees]C for 20 min. After sterilization, the bags were allowed to cool for 24 hrs. and inoculated aseptically in the laminar airflow cabinet with actively growing mycelium of the pure culture of Pleurotus florida from PDA slant. A piece of pure culture (1 x 1cm) was placed aseptically into the mouth of each bag using sterile scalpel and forceps. After inoculation, the bags were incubated at 25 [+ or -] 1[degrees]C under dark condition to allow the full mycelial ramification of the grains. After about one week, the bags were shaken to spread the mycelium evenly through the bags.
Cultivation of the Mushroom:
Preparation of substrates:
Date palm wastes and sawdust were used as basal substrates. Whole date palm wastes collected from the Ministry of Agriculture, Riyadh City, Saudi Arabia, were chopped into small pieces (2-3cm). The substrates were soaked overnight in water and excess water was drained off. After soaking, the substrates (50-60% moisture content) were separately subjected to short composting (1week).
Preparation of the mushroom spawn bags:
The composted substrates (7 parts of date palm wastes and 3 parts of sawdust on the dry weight basis)) and 10% of wheat bran supplement as a source of nitrogen were thoroughly mixed and finally maintained at 70% moisture content [27,8]. The mixture was divided into batches of 600 g each and packed into heat resistant polypropylene bags (30 x 14 cm). The openings of the bags were plugged with cotton and bind with rubber band. The filled bags were pasteurized using steam for 5 hrs. at 60-80[degrees]C in a closed chamber and then these bags were left inside the steam chamber for 24 hrs to cool down. The pasteurized cooled bags were inoculated aseptically with spawn at the rate of 2.0% of the wet weight basis of substrate (12g/ bag ([approximately equal to] 70 g/kg dry substrate as recommended by Ahmed et al, ). The inoculated bags were placed inside a spawn running room under dark conditions. During spawn running, the temperature was controlled between 22-26 [degrees]C. After 4 weeks of incubation, bags were fully covered with white mycelia. Inoculated bags were transferred to the growing room and opened by removing the cotton plug and rubber band from the mouth of each bag.
The conditions of the growing room were adjusted where the temperature and relative humidity were maintained between 22-25[degrees]C and 80-95%, respectively with sufficient light and ventilation for the development of normal fruiting bodies by means of humidifier and cooler and by sprinkling water on walls and floor. The moisture requirements of the bags were accomplished by sprinkling water on them twice a day (in the morning and evening) using sprinkler. Water spraying was done until the mushroom was matured enough to be harvested. Mushroom was harvested as soon as the fruiting bodies developed and attained their full size above the substrate. After each flush, two holes opposite to each other of the upper position of bags were opened with a blade by removing the plastic sheet in "D" shape. A small layer of substrate was scrapped off from all the side of the beds after each harvest. Mushroom was harvested in flushes over a 45-day period.
Harvesting of the Mushroom:
Harvesting of mushroom was conducted at four different flushes. During these flushes the mushroom was harvested based on the maturation and color appearance of the fruiting bodies.
For the productivity evaluation, the following parameters were used: total number of flushes, yield of each bag and total mushroom yield (g) (fresh and dry weight), moisture content (%), biological efficiency (%) and bioconversion efficiency (%).
The total yield was estimated on the basis of 1kg dry substrate by measuring the fresh and dry weight. The total yield was recorded by adding the fresh as well as dry weight of mushroom of all flushes. The fresh (wet) weight of mushroom was recorded after harvesting of each flush. The dry weight of mushroom (g) was determined after drying the fruiting bodies.
The moisture content of mushroom was determined after drying the fruiting bodies in an air oven at 67 [degrees]C for 24 hrs. . It was expressed in percent and calculated by the formula:
Moisture content (%) = [Fresh weight of mushroom (g)--Dry weight of mushroom (g)/Fresh weight of mushroom (g)] x 100
The biological efficiency (BE) (yield of fresh mushroom per kg substrate on dry weight basis) was calculated by the formula recommended by Chang et al.  as:
BE (%) = [Fresh weight of mushroom (g) / Dry weight of substrate (g)] x 100
The bioconversion efficiency (BCE) (yield of dry mushroom per kg substrate on dry weight basis) was calculated by the formula recommended by Elenwo and Okere  as:
BCE (%) =[Dry weight of mushroom (g) / Dry weight of substrate (g)] x 100
RESULTS AND DISCUSSIONS
Commercial production of oyster mushrooms is largely determined by the availability and utilization of cheap materials of which agricultural lingo-cellulosic waste represents the ideal and most promising substrates for cultivation. The substrates used in this study can be considered practical and economically feasible due to their availability throughout the year at little or no cost in large quantities. Utilization of these agro-wastes for the production of oyster mushrooms could be more economically and ecologically practical as mentioned by Ahmed et al.  and Ingale and Ramteke .
Number of flushes:
The obtained results proved that the oyster mushroom (Pleurotus florida) gave maximum (4 flushes) by using 70g spawn rate per kg substrate dry weight basis. Similar results were found by Bhatti et al.  by using 70g spawn rate per kg on wheat straw dry weight basis with the oyster mushroom (Pleurotus ostreatus). Several investigators stated that 3 flushes were observed [24,23,37,47,1,8]. The time required for the flushes was 9, 19, 31 and 45 days for the 1st, 2nd, 3rd and 4th flush, respectively from the opening of bags.
The harvest yield of Pleurotus florida was estimated on the basis of 1kg dry substrate by measuring the fresh and dry weight. The yield of mushroom from the 4 flushes is shown in Figure 1. The fresh weight was 197.82, 156.71, 105.8 and 76.31g/kg dry substrate, whereas the dry weight was 24.6, 19.34, 16.64 and 11.02 g/kg dry substrate in case of 1st, 2nd, 3rd and 4th flush, respectively. It was observed that, the harvest yield decreased with increasing flushing period. Similar trend was observed by several investigators [23,37,1]. This finding was in agreement with the finding of Shah et al.,  who stated that the maximum yield was observed in the first flush.
The total yield of Pleurotus florida in terms of fresh weight was 536.64 g/kg dry substrate and dry weight was 71.6 g/kg dry substrate (Figure 2). Similar result was found by Ingale and Ramteke  who observed that the total yield of Pleurotus florida in terms of fresh weight was 566.42 g/kg dry substrate and dry weight was 79.46 g/kg dry substrate (soybean straw). Also, Mintesnot et al.  observed that the total fresh weight ranged from 377 to 665g/kg dry substrate in case of Pleurotus florida on different substrates. In another study, the total fresh weight ranged from 723.66 to 875.66g/kg dry substrate in case of Pleurotus florida on different substrates . The obtained result was higher than that found by Kumari and Achal  who stated that the maximum fresh weight (29.27 g/kg substrate) as well as dry weight (7.32 g/kg substrate) was found when Pleurotus ostreatus was grown on wheat straw as substrate. Generally, the yield of the mushroom is directly related to the spread of the mycelium into the substrates .
Moisture content of the fruiting bodies:
The moisture content (%) of fruiting bodies of mushroom from the 4 flushes is shown in Figure 3. The moisture content was 87.32, 87.71, 84.23 and 85.92 % in the 1st, 2nd, 3rd and 4th flush, respectively. It is noticed that the moisture content of fruiting bodies in the first two flushes was higher than the others and generally the difference between the flushes was very small. On the other hand, Abeer et al.  observed that the moisture content of fruiting bodies decreased with increasing flushing periods.
The biological efficiency (BE) was calculated to determine how the mushrooms utilized efficiently the nutrients present in the substrates as mentioned by Oseni et al. (2012). The biological efficiency obtained from the 4 flushes is presented in Figure 4. It was 19.78, 15.67, 10.58 and 7.63% in the 1st, 2nd, 3rd and 4th flush, respectively. It is noticed that, the biological efficiency decreased with increasing flushing period. Similar trend was observed by Abeer et al.  who reported that the biological efficiency decreased from 78.94% after 1st flushing period to 31.66 after 3rd flushing period. In the present study, the total biological efficiency was 53.66% (Figure 5). Similar result was found by Ingale and Ramteke  who observed that the total biological efficiency was 56.64 % in case of Pleurotus florida on soybean straw. The obtained result was approximately similar to the findings of some researchers. Mintesnot et al.  observed that the biological efficiency ranged from 37.68% to 66.30% in case of Pleurotus florida on different substrates, whereas Alam et al.  observed that the biological efficiency ranged from 45.21% to 125.70% in case of Pleurotus ostreaus on sawdust and rice straw. In another study by Ahmed et al. , the biological efficiency ranged from 72.36% to 87.56% in case of Pleurotus florida on different substrates. On the other hand, the obtained result was higher than the findings of Elenwo and Okere  who mentioned that the biological efficiencies obtained from Pleurotus tuber-regium on corn cob, Pleurotus osteratus var florida on corn husk and Volvariella volvacea on corn husk were 5.66%, 2.44%, and 0.8%, respectively. They stated also that the low biological efficiency obtained could be attributed to the short production cycle. Generally, as mentioned by Mane et al. ) cultivation of the oyster mushroom on similar by-products manifested variable levels of BE. These variations are mainly related to spawn rate, fungal species used and supplement added to the substrate.
The bioconversion efficiency (BCE) obtained from the 4 flushes is presented in Figure 4. It was 2.46, 1.93, 1.66 and 1.10% in case of 1st, 2nd, 3rd and 4th flush, respectively. It decreased also with increasing flushing period. The total bioconversion efficiency was 7.15% (Figure 5). Similar result was found by Ingale and Ramteke  who observed that the bioconversion efficiency was 7.94%in case of Pleurotus florida on soybean straw. The obtained result was higher than the findings of Elenwo and Okere  who mentioned that the bioconversion efficiency of 0.67%, 0.36% and 0.09% was obtained from Pleurotus tuber-regium on corn cob, Pleurotus ostreatus var florida on corn husk and Volvariella volvacea on corn husk, respectively. They reported that the low bioconversion efficiency is in agreement with the low yield obtained due to a relatively short production cycle.
Generally, the difference between the results from this investigation with the results reported by other research workers may be due to the variation in controlled, semi controlled conditions, physiological requirements for cultivation of oyster mushroom e.g., constant temperature, humidity and light arrangements .
Based on the result of current study, it could be concluded that the adjustment of environmental conditions (favorable temperature, relative humidity, moisture content) is very important to enhance production of fruiting bodies of the oyster mushroom (Pleurotus florida), especially in Saudi Arabia. If country's environmental conditions are not suitable to provide this temperature and relative humidity, mushroom should be cultivated in controlled conditions. It is evident that, Pleurotus florida is a suitable species for cultivation on the mixture of date palm wastes and sawdust in case of productivity and can be cultivated under environmentally controlled conditions. This will ensure sustainable production which will result in a constant supply of mushroom in the market.
Received 12 November 2014
Received in revised form 31 December 2014
Accepted 22 January 2015
Available online 25 February 2015
The authors extend their appreciation to the Deanship of Scientific Research at King Saud University for funding the work through the research group project No. RGP-VPP-134.
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(1) Fahad Alkoaik, (1) Ahmed Khalil, (1) Ronnel Fulleros and (2) Renato G. Reyes
(1) Department of Agricultural Engineering, College of Food and Agriculture Sciences, King Saud University, Saudi Arabia.
(2) Center for Tropical Mushroom Research & Development Center, Central Luzon State University, Science City of Munoz, Nueva Ecija, Philippines.
Corresponding Author: Fahad Alkoaik, Department of Agricultural Engineering, College of Food and Agriculture Sciences, King Saud University, Saudi Arabia.
Tel: +966 11 504141754, Fax: +966 11 4678502, E-mail: firstname.lastname@example.org
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|Author:||Alkoaik, Fahad; Khalil, Ahmed; Fulleros, Ronnel; Reyes, Renato G.|
|Publication:||Advances in Environmental Biology|
|Date:||Feb 1, 2015|
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