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Cultivation of oyster mushroom (Pleurotus florida) on date palm was and their mixture was .... During spawn running, the temperature was controlled between ...
Advances ces in Environmental Biology, 9(14) July 2015, Pages: 207-215

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Advances ces in Environmental Biology ISSN-1995-0756

EISSN-1998-1066

Journal home page: http://www.aensiweb.com/AEB/

Preliminary Study for Cultivation of Oyster Mushroom (Pleurotus (Pleurotus florida) on Agricultural Wastes in Saudi Arabia 1,2Ahmed 1 2

Ismail Khalil, 1Fahad Nasser Alkoaik and 1Ronnel Blanqueza Fulleros

Department of Agricultural Engineering, College of Food and Agriculture Agriculture Sciences, King Saud University, Saudi Arabia Department of Environmental Studies, Institute of Graduate Studies and Research, University of Alexandria, Alexandria, Egypt

ARTICLE INFO Article history: Received 28 May 2015 Accepted 10 July 2015 Available online 15 July 2015 Keywords: Agricultural wastes, mushroom cultivation, Pleurotusflorida

ABSTRACT Cultivation of oyster mushroom (Pleurotus florida) on date palm wastes and sawdust and their mixture was attempted primarily to determine the possibility pos of its cultivation under environmentally controlled conditions in Saudi Arabia.After Arabia. soaking overnight, the substrates were separately subjected to short composting. The composted substrates (date palm wastes, sawdust and their mixture (7 parts of date palm wastes and 3 parts of sawdust on the dry weight basis) were maintained at 70% moisture content and packed into heat resistant polypropylene bags. Then, the bags were pasteurized, cooled and inoculated with grain spawn. After completion of spawn running, the matured spawn bags were transferred to the growing room. The obtained results showed that theoyster mushroom (P. florida) yielded maximum flushes (5, 3 and 4 flushes) on date palm wastes, sawdust and their mixture, respectively. The time required requi for the flushes varied according to the substrate used. The total fresh weight was 255.3, 255.14 and 331.1g/kg dry substrate and the total dry weight was 39.37, 39.82 and 51.18 g/kg dry substrate in case of date palm wastes, sawdust and their mixture, respectively. The moisture content (%) of fruiting bodies varied also according to the substrate and to each flush. The total biological efficiency was 25.54, 25.51 and 33.11% and bioconversion efficiency was 3.93, 3.98 and 5.13% in case of date palm wastes, s, sawdust and their mixture, respectively. It could be concluded that P.florida can be cultivated under controlled conditions.The The performance of the mixture was evident by the elevated yield, biological efficiency and bioconversion efficiency values.

© 2015 AENSI Publisher All rights reserved. reserved To Cite This Article: Ahmed Ismail Khalil, Fahad Nasser Alkoaik and RonnelBlanqueza Fulleros., Preliminary Study for Cultivation of Oyster Mushroom (Pleurotus florida)on )on Agricultural Wastes in Saudi Arabia. Arabia Adv. Environ. Biol., 9(14), 207-215 215, 2015

INTRODUCTION Vast quantities of agricultural and agro-industrial agro industrial residues that are generated as a result of diverse practices represent one of the most important energy-rich energy resources available [10].. Accumulation of this biomass in large quantities results not only in environmental deterioration, but also in the loss of potentially valuable resources [10,26] which can be processed to yield food, feed and a variety of chemicals [10]. So, it is necessary to find new technologies chnologies to overcome this issue [36]. The bioconversion of agriculture and industrial wastes into food has attracted the world attention in recent years. There is greater demand for the available world supplies of food and second are the prices of those thos foods that are increasingly steeply [4,20,23]. The cultivation of mushrooms has a great potential for the production of protein rich quality food and for recycling of cellulosic agro-residues agro residues and other wastes [37] and serves as the most efficient and economically nomically viable biotechnology for the conversion of lignocellulose waste materials to high quality food and this will naturally open up new job opportunities especially in rural areas [19,22]. Mushroom cultivation, apart from being a source of food production, production, can be a means of livelihood and a source of economic empowerment for women in both urban and rural areas, and for small holder farmers [32]. This technology can also limit air pollution associated with burning agriculture wastes as well as to decrease decrea environmental pollution due to unutilized agricultural wastes [1]. Mushroom cultivation is a useful method of environmental waste management and waste disposal. Many agricultural and industrial by-products by can find uses in mushroom production[3].

Corresponding Author: Ahmed Ismail Khalil, Department of Agricultural Engineering, College of Food and Agriculture Sciences, King Saud University, Saudi Arabia. E-mail: [email protected]

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Mushrooms are fungi that belong to Basidiomycotina or Ascomycotina and they produce lignocellulosic enzymes, which degrade complex organic matter and absorb the soluble substances [13]. Many types of mushrooms both edible and non-edible exist. The edible mushrooms are widely used as human food [12]. The cultivation of edible mushrooms offers one of the most feasible and economic methods for the bioconversion of agro-lignocellulosic wastes [7,16]. Oyster mushrooms are one of the most popular edible mushrooms and belong to the genus Pleurotus and the family Pleurotaceae[27]. The common name “oyster mushroom” refers to several species of edible mushroom belonging to the genus Pleurotus[34] and comes from the white shell-like appearance of the fruiting body [38]. Oyster mushrooms are ideally suitable for cultivation under both temperate and tropical climatic conditions [5]. Cultivation of oyster mushrooms is becoming popular throughout the world because of their abilities to grow at a wide range of temperatures and to utilize various lignocelluloses. Pleurotus species have extensive enzyme systems capable of utilizing complex organic compounds that occur as agricultural wastes and industrial by-products. These mushrooms are also found to be one of the most efficient lignocelluloses solid state decomposing types of white rot fungi [8]. The genus Pleurotus, is widely cultivated due to its favorable organoleptic and medicinal properties, simple and low cost production technology and higher biological efficiency [15]. Generally, the cultivation of oyster mushroom can play an important role in managing organic wastes whose disposal has become a problem as mentioned by Das and Mukherjee [17]. Oyster mushroom (Pleurotus florida) is an edible mushroom having excellent flavor and taste [31] and is increasingly becoming popular as protein-rich delicious vegetable [33]. Pleurotus florida cultivation is popular due to low cost technology and easy availability of various substrates for its cultivation[11]. It is widespread in temperate, subtropical and tropical zones and it shows the highest yield among the Pleurotus species [27]. Therefore, the objective of this study was to evaluate the cultivation of oyster mushroom (Pleurotus florida) on some common and abundantly available agricultural wastes in Saudi Arabia as a preliminary study. 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, during the period of February to June 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 4ºC and recultured bimonthly. Preparation of Spawn: Wheat grains were boiled in water bath for 10-15min at the ratio of 1:1 (wheat grains: water) and allowed to cool for 15 min, then the water was drained and the boiled grains were dried on cotton cloth. Then, the grains were packed tightly in heat resistant polypropylene bags (30 x 14 cm). One bag contained 350g of the prepared grains.The bags were plugged with cotton and covered with aluminum foil. The packed bags were sterilized in an autoclave at 121ºC 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 (1x1cm) was placed aseptically into the mouth of each bag using sterile scalpel and forceps. After inoculation, the bags were incubated at 25 ± 1ºC for 3 weeks under dark condition to allow the full mycelialramification 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 as agricultural wastes were used as basal substrates. Dried date palm wastes (branches and leaves) collected from the Educational Farm, Department of Agricultural Engineering, College of Food and Agriculture Sciences, University of King Saud, Riyadh City, Saudi Arabia, were chopped into small pieces (2-3 cm). Sawdust was collected from wood market, Riyadh City, Saudi Arabia. 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 (date palm wastes, sawdust and their mixture (7 parts of date palm wastes and 3 parts of sawdust on the dry weight basis)) were finally maintained at 70% moisture content [6,28]. Then, the substrates were packed in heat resistant polypropylene bags (30 ×14 cm) and the bags were plugged with cotton and bind with rubber band. Each bag contained 600g wet substrate. Each treatment was replicated three times.

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The filled bags were pasteurized using steam for 5 hrs at 60-80oC in a closed chamber and then these bags were kept 24 hrs for cooling. The pasteurized cooled bags were inoculated aseptically with spawn at the rate of 2.0% of the wet weight basis of substrate (12 g/ bag (≈70 g/kg dry substrate)) [1]. The inoculated bags were placed inside a spawn running room under dark conditions. During spawn running, the temperature was controlled between 22-26oC for spawn running. After completion of spawn running (4 weeks, when the bags were fully covered with white mycelia), the bags were transferred to the growing room and opened by remove the cotton plug and rubber band from the mouth of each bag. Temperature, humidity and light are basic factors for the proper growth and development of mushroom. Efforts were made to maintain these requirements by furnishing mushroom growing room.The conditions of the growing room were adjusted but not reach to the optimum, where the temperature and relativehumidity were maintained between 21-25ºC and 50-95%, respectively with little light and ventilationfor the development of normal fruiting bodies. The relative humidity was maintained between 50-95% by using 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 47-day period. Harvesting of the Mushroom: Harvesting of mushroom was conducted at different flushes according to the substrate. During these flushes the mushroom was harvested based on the maturation and color appearance of the fruiting bodies. Productivity Evaluation: 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 ºC for 24 hrs[41]. 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. [14] and Chang and Miles[13] 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[18] as: BCE (%) =[Dry weight of mushroom (g) / Dry weight of substrate (g)] x 100 RESULTS AND DISCUSSION In the present study two agricultural wastes (date palm wastes and sawdust) and their mixture were used for cultivation of the oyster mushroom (Pleurotus florida). The substrates used 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. [1] andIngale and Ramteke[23]. Number of flushes: The obtained results proved that Pleurotus florida gave maximum flushes (5, 3 and 4 flushes)on date palm wastes, sawdust and their mixture, respectively) by using 70g spawn rate per kg substrate dry weight basis. Similar results were found by Bhattiet al. [9] 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 [1,6,21,23-25,31,35,40]. The time required for the flushes from the opening of bagsvaried according to the substrate used. In case of date palm wastes, it was 8, 23, 31, 38 and 47 days for the 1st, 2nd, 3rd, 4thand 5thflush, respectively. In case of sawdust, it was 8, 38 and 47 days for the 1st, 2nd and 3rd flush, respectively, whereas in case of the mixture, it was 8, 23, 38 and 47 days for the 1st, 2nd, 3rd and 4th flush, respectively. Harvest yield: 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 flushes is shown in Figure 1. In case of date palm wastes,

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the fresh weight was 82.15, 20.28, 49.72, 67.85 and 35.3g/kg dry substrate, whereas the dry weight was 15.94, 5.7, 5.89,6.43 and 5.41g/kg dry substrate in the 1st, 2nd, 3rd, 4thand 5thflush, respectively. In case of sawdust, the fresh weight was 67.93, 98.15 and 89.06 g/kg dry substrate, whereas the dry weight was 22.5, 8.37 and 8.95g/kg dry substrate in the 1st, 2nd and 3rd flush, respectively, whereas in case of the mixture, the fresh weight was 151.55, 44.56, 98.7 and 36.28 g/kg dry substrate, whereas the dry weight was 26.61, 12.96, 7.15 and 4.46g/kg dry substrate in the 1st, 2nd, 3rd and 4thflush, respectively.The harvest yield varied due to effect of different substrates at different flushes. The results presented in Figure 2 indicate the total yield of Pleurotus florida in terms of fresh weight was 255.3, 255.14 and 331.1g/kg dry substrate and dry weight was 39.37, 39.82 and 51.18 g/kg dry substrate in case of date palm wastes, sawdust and their mixture, respectively. The harvest yield on the mixture was higher than that found on each separate substrate. Our results are closerto those found by Khan et al. [25] who observed that the total fresh yield of Pleurotus (flabellatus) djamor (R-22) was 233g/kg substrate (mixed sawdust from different woods). The obtained results were higher than those found by Kumari and Achal [28] 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. On the other hand, the obtained results were lower than those found by other investigators. Ingale and Ramteke[23] 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. [30] 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 by Ahmed et al. [1] the total fresh weight ranged from 723.66 to 875.66g/kg dry substrate in case of Pleurotus florida on different substrates. In the present study, the yield of mushroom was affected by different substrates. The highest total yield of mushroom was recorded on the mixture followed by yield on date palm wastes and the lowest yield was found on sawdust. Generally, the yield of the mushroom is directly related to the spread of the mycelium into the substrates [39]. The variations observed in yield may, therefore be attributed to the complexity of substrates in terms of their cellulose content resulting from the difference in the rate of degradation by the mushroom enzymes[35].

Fig. 1: Changes in wet and dry weight (g) of oyster mushroom (Pleurotusflorida)during the cultivation on date palm wastes (A), sawdust (B) and their mixture (C). Values are means of 3 replicates ± standard deviations. Moisture content of the fruiting bodies: The moisture content (%) of fruiting bodies of mushroom from the flushes is shown in Figure 3. In case of date palm wastes, it was 80.42, 57.71, 88.21, 90.56 and 84.61% in the 1st, 2nd, 3rd, 4thand 5th flush, respectively. In case of sawdust, it was 61.97, 91.57 and 89.86% in the 1st, 2nd and 3rd flush, respectively, whereas in case of the mixture, it was 82.41, 61.77, 92.76 and 86.43% in the 1st, 2nd, 3rd and 4th flush, respectively. The moisture content of fruiting bodies varied due to effect of different substrates at different flushes. Generally, there was slight variation indicating that moisture content is independent of the substrate as mentioned by Ahmed et al.

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[1]. On the other hand, Hashem et al. [21] observed that the moisture content of fruiting bodies decreased with increasing flushing periods.

Fig. 2: Total weight (g) of oyster mushroom (Pleurotus florida) during the cultivation on date palm wastes, sawdust and their mixture. Values are means of 3 replicates ± standard deviations. (DP: date palm, SD: sawdust).

Fig. 3: Changes in moisture content (%) of oyster mushroom (Pleurotus florida) during the cultivation on date palm wastes (A), sawdust (B) and their mixture (C). Values are means of 3 replicates ± standard deviations. Biological efficiency: The biological efficiency (BE) is indicating the conversion of substrate mass to mushroom fruiting bodies [40]. The BE was calculated to determine how the mushrooms utilized nutrients present in the substrates efficiently [35]. The BE obtained from the flushes is presented in Figure 4. In case of date palm wastes, it was 8.22, 2.03, 4.97, 6.79 and 3.53% in the 1st, 2nd, 3rd, 4th and 5th flush, respectively. In case of sawdust, it was 6.79, 9.81 and 8.91% in the 1st, 2nd and 3rd flush, respectively, whereas in case of the mixture, it was 15.15, 4.46, 9.87 and 3.63% in the 1st, 2nd, 3rd and 4th flush, respectively. It is noticed that, the biological efficiency varied due to effect of different substrates at different flushes.The total BEwas 25.54, 25.51 and 33.11% in case of date palm wastes, sawdust and their mixture, respectively (Figure 5). The highest value was found on the mixture,

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followed by date palm wastes and then by sawdust. The obtained results were higher than the findings of Elenwo and Okere[18] who mentioned that the BE obtained from Pleurotus tuber-regiumon corn cob, Pleurotus osteratus var florida on corn huskand Volvariellavolvaceaon corn huskwas 5.66%, 2.44%, and 0.8%, respectively. They stated also that the low BE obtained could be attributed to the short production cycle. The obtained resultswere approximately similar to the findings of Mintesnot et al. [30] who observed that the BE ranged from 37.68% to 66.30% in case of Pleurotusflorida on different substrates.On the other hand, the obtained results were lower than the findings ofAlam et al. [2] who observed that the BE ranged from 45.21% to 125.70% in case of Pleurotus ostreaus on sawdust and rice straw. Also, Ahmed et al. [1] stated that the BE ranged from 72.36% to 87.56% in case of Pleurotus florida on different substrates. Generally, as mentioned by Mane et al.[29], 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.

Fig. 4: Changes in biological efficiency (%) of oyster mushroom (Pleurotus florida) during the cultivation on date palm wastes (A), sawdust (B) and their mixture (C). Values are means of 3 replicates ± standard deviations.

Fig. 5: Total biological efficiency (%) of oyster mushroom (Pleurotus florida) during the cultivation on date palm wastes, sawdust and their mixture. Values are means of 3 replicates ± standard deviations. (DP: date palm, SD: sawdust).

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Bioconversion efficiency: The bioconversion efficiency (BCE) obtained from the flushes is presented in Figure 6. In case of date palm wastes, it was 1.59, 0.57, 0.59, 0.64 and 0.54% in the 1st, 2nd, 3rd, 4th and 5th flush, respectively. In case of sawdust, it was 2.25, 0.84 and 0.89% in the 1st, 2nd and 3rd flush, respectively, whereas in case of the mixture, it was 2.66, 1.3, 0.72 and 0.45% in the 1st, 2nd, 3rd and 4th flush, respectively. It is noticed that, the BCE varied also due to effect of different substrates at different flushes. The BCE decreased with increasing flushing period; especially in case of the mixture. The total BCE was 3.93, 3.98 and 5.13% in case of date palm wastes, sawdust and their mixture, respectively (Figure 7).The highest value was found on the mixture, followed by date palm wastes and then by sawdust. The obtained results were higher than the findings of Elenwo and Okere[18] who mentioned that theBCE of 0.67%, 0.36% and 0.09% was obtained from Pleurotus tuber-regiumon corn cob, Pleurotusostreatusvarfloridaon corn husk andVolvariellavolvaceaoncorn husk, respectively. They reported that the low BCE is in agreement with the low yield obtained due to a relatively short production cycle. Closer result was found by Ingale and Ramteke[23] who observed that the BCE was 7.94%in case of

Fig. 6: Changes in bioconversion efficiency (%) of oyster mushroom (Pleurotus florida)during the cultivation on date palm wastes (A), sawdust (B) and their mixture (C). Values are means of 3 replicates ± standard deviations.

Fig. 7: Total bioconversion efficiency (%) of oyster mushroom (Pleurotus florida) during the cultivation date palm wastes, sawdust and their mixture. Values are means of 3 replicates ± standard deviations. (DP: date palm, SD: sawdust).

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Pleurotusfloridaonsoybean straw: The lowharvest yield,BE and BCE obtained from this study however could be attributed to the growth conditions (temperature, humidity and light), where it is the first time to cultivate this mushroom in our laboratories. Efforts were made to adjust these conditions but not reach to the optimum, where the temperature and relative humidity were 21-25ºC and 50-95%, respectively with little light and ventilation for the development of fruiting bodies.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. The response of different substrates also show differences in respect of time taken for maturation of fruiting bodies, period between flushes, number of flushes and yield as mentioned by Bhatti et al. [9]. Conclusion: From the obtained results, it can be concluded that the growth and yield of Pleurotus florida varied widely, depending on the kind of substrate used. It is evident that, Pleurotus floridais the suitable species for cultivation on date palm wastes and sawdust and their mixture in case of productivity and can be cultivated under environmentally controlled conditions. The adjustment of environmental conditions (temperature, relative humidity, moisture content) is very important to enhance production of fruiting bodies of Pleurotus florida; especially in Saudi Arabia. The performance of the mixture of date palm wastes and sawdust was evident by the elevated yield, BE and BCE values. This will enable the farmer to get extra income and more protein harvest from his agricultural waste.Also, this will ensure sustainable production which will result in a constant supply of mushroom in the market. ACKNOWLEDGMENTS 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. REFERENCES [1] Ahmed, S.A., J.A. Kadam, V.P. Mane, S.S. Patil and M.M.V. Baig, 2009. Biological efficiency and nutritional contents of Pleurotus florida (Mont.) Singer cultivated on different agro-wastes.Nature Sci., 7(1): 44-48. [2] Alam, N., S.M.R. Amin and N.C. Sarker, 2007. Efficacy of five different growth regulators on the yield and yield contributing attributes of Pleurotusostreatus(Jacquin ex Fr.) Kummer. Bang. J. Mush., 1 (1): 51-55. [3] Alemu, F., 2014. Cultivation of oyster mushroom (Pleurotusostreatus) on teff straw (by-product of agriculture) at Dilla University, Ethiopia.Global Adv. Res. J. Med. Med. Sci., 3(8): 205-215. [4] Ali, M. A., J. Ashraf, N. A. Khan and A.Rehman, 2012. Amendment of cotton waste with different waste materials of sugarcane industry for mycelial growth and fructification of oyster mushroom.Pak. J. Phytopathol.,24(1): 6-10. [5] Amin, S.M.R., C.S. Nirod, M. Moonmoon, J. Khandaker and M. Rahman, 2007. Officer’s Training Manual. National Mushroom Development and Extension Centre, Savar, Dhaka, Bangladesh. pp. 7-17. [6] Ashraf, J., M. A. Ali, W. Ahmad, C. M. Ayyub and J. Shafi, 2013. Effect of different substrate supplements on oyster mushroom (Pleurotusspp.) production.Food Sci. Technol., 1(3): 44-51. [7] Bano, Z., M.N. Shasirekha and S. Rajarathnam, 1993. Improvement of the bioconversion and biotransformation efficiencies of the oyster mushroom (Pleurotussajor-caju) by supplementation of its rice straw with oil seed cakes.Enzyme Microb. Technol., 15: 985-989. [8] Baysal, E., H. Peker, M. Kemal and A. Temiz, 2003. Cultivation of oyster mushroom on waster paper with some added supplementary materials. Bioresour. Technol., 89: 95-97. [9] Bhatti, M.I., M.M. Jiskani, K.H. Wagan, M.A. Pathan and M.R. Magsi, 2007. Growth, development and yield of oyster mushroom, Pleurotusostreatus (Jacq. Ex.Fe.) kummer as affected by different spawn rates. Pak. J. Bot., 39(7): 2685-2692. [10] Bisaria, V.S., 1991. Bioprocessing of agro-residues to glucose and chemicals.In: Martin, A.M. (Ed.), Bioconversion of Waste Materials to Industrial Products. Elsevier Applied Science, London and New York, pp.187-223. [11] Biswas, S., M.S. Hoque and K.U. Ahmed, 2009. Effect of a mineral supplement on growth, yield and nutritional status of oyster mushroom (Pleurotusostreatus).Bang. J. Mush., 3(2): 51-58. [12] Chang, S.T., 1980. Mushroom production in South East Asia. Mushroom Newslett. Trop., 1(4): 5-10. [13] Chang, S.T. and P.G. Miles, 1989. Edible Mushrooms and Their Cultivation.1st edition.CRC Press, Boca Raton, Florida, USA. [14] Chang, S.T., O.W. Lau and K.Y. Cho, 1981.The cultivation and nutritive value of Pleurotussojar –caju. European J. Appl. Microbiol.Biotechnol.,12: 58-62.

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