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Greener Journal of Agricultural Sciences

Vol. 8(6), pp. 110-118, June, 2018

ISSN: 2276-7770; ICV: 6.15

Copyright ©2018, the copyright of this article is retained by the author(s)

DOI Link: http://doi.org/10.15580/GJAS.2018.6.051518071

http://gjournals.org/GJAS

 

 

 

 

Evaluation of Grevillae Robusta Dry Leaves Supplemented with Cotton Seed Waste for the Cultivation of Oyster mushroom (Pleurotusostreatus)

 

 

ASEFA Keneni

 

Greener Journal of Agricultural Sciences, vol. 8, no. 6, pp. 110-118, June 2018

 

 

Department of Biology, Ambo University, Ambo, P.O.Box.19, Ethiopia.

 

 

 

 

ARTICLE INFO

ABSTRACT

 

Article No.: 051518071

Type: Research

DOI: 10.15580/GJAS.2018.6.051518071

 

 

At present more emphasis has been given to mushroom production for the nutritionand, medicinaluses and waste recycling technology. The main aim of the present study was to report the usability of Greviillaerobusta dry foliage as a major substrate for production of oyster (Pleurotusostreatus) mushroom with supplement of different proportion of cotton seed waste. The culture of the oyster mushroom was maintained on potato dextrose agar, and the spawn was prepared on yellow colored sorghum and sterilized substrate was inoculated with 5% of the spawn wetbasis on dry basis of the substrate.The experimental design constitutes nine treatments (T1-T9) in three replicates from the middle of November2017 to the end of February 2018. Fastest mycelia run was observed in the treatments T9, T7 and T8: 10 days each from inoculation, while the slowest mycelia runwas observed in treatment T5 and T6: 19 days each from inoculation. Longest production cycle wasobserved for treatment T3, T5 and T6: 111days each, while the shortest production cycle was recordedfor treatment 8: 100days. Highest fresh weight1246.5 g /600g dry substrate was recorded for T 9; highest number (130)of fruits recorded for T4 while largest cap diameter(8cm) was recorded for treatment T6.The T1 showed the  lowest fresh weight, 538g/600 g/dry weight of the substrate, with  70 fruits and 6.5cm cap diameter.No Significantdifference were observed for the stipe length of the different treatments. Highest biological efficiency was recorded for T9: 208; and 89.5 for T1.Even though the cotton seed waste proportion supplementation varied for the treatments all except T1 gave more than 100% biological efficiency which makes Grevillae dry leaves, a good substratum for mass production of oyster mushroom which can produce good quality mushroom fruit bodies.

 

Submitted: 15/05/2018

Accepted:  30/05/2018

Published: 15/06/2018

 

*Corresponding Author

ASEFA Keneni

E-mail: asefa_keneni

@yahoo.com

 

 

Keywords:

Cotton seed waste, growth, Grevlliae dry leave, oyster mushroom, yield

 

 

 

 

 

 

INTRODUCTION

 

Mushrooms are basically fungi which have a flashy and spore bearing fruiting body growing in fields, forests; on manure heaps, on water channels, on hilly areas, on damp rotten log of wood trunks of trees, on decomposing organic matter, in damp soil rich in organic substances, mostly during and just after rainy season.They comprise a large heterogeneous group having various shapes, sizes and color, all quite different in character, appurtenance and edibility. Based on standard morphology the word “mushroom was mostly used to describe those fungi that have stem (stipe), a cap (pileus), and gills (lamellae)or the pores on the under sides of the cap. Only a fruiting body of the mushroom can be seen whereas the rest of the mushroom remains underground as mycelium (Chang and Miles 1992).

Mushroom has been recognized as alternative sources of good quality protein. They are capable of producing the highest quality of protein per unit area and time, from the worthless agro waste which are available in huge amount all-round the year and they, represents as one of greatest untapped resources of nutrition and palatable food of the future. The nutritional value of the edible mushroom can be compared with eggs, milk, and meat (Dabaet al., 2008). And the mushroom contents of essential amino acids are high and close to the need of the human body. It is easily digestible and it has no cholesterol (Chang and Buswell, 1996).

Mushroom with protein content ranging from 3-7% when fresh to 25-40% when dry can play an important role in enriching human diet when meat sources are limited. The mushroom protein content is almost equal to that of corn, milk, and legumes, although still lower than meat , fish and egg (Sparks,2006).Mushrooms are considered as a functional food, which can provide health benefits beyond the traditional nutrients they contain (Miles and Chang, 2004). Without doubt, edible mushroom in fresh, cooked or processed forms are nutritionally sound, tasteful food source for most people and can be a significant dietary component for vegetarians (Daba et al., 2008). From a number of cultivated mushrooms, oyster mushroom (Pleurotusostreatus) is the most preferred species due to its ability to utilize different organic waste due the presence of efficient enzymes such as cellulose, hemicelluloses, and lignin, for fast growth.

Several species of the genus Pleurotus have great commercial value in the global market of edible cultivated mushrooms, which reportedly grew from 2.13 million tons in 1996 to 3.43 million tons in 2007 (Okuda et al., 2009).Nutritional value of oyster mushroom is also appreciable. 3.5 oz. of oyster mushroomprovides 38 Calories and contain, 15%-25% of Protein, 2.2g of Fat, 6.5g of Carbohydrate, 2.8g of Fiber, and also contains Vitamin B1 (Thiamine) 0.56 mg, Vitamin B2 (Riboflavin) 0.55 mg, Vitamin B3 (Niacin) 12.2 mg, Phosphorus 140 mg, Calcium 28 mg, Iron 1.7 mg3. Comparatively it is high in fiber and low in carbohydrate and fat which make it more potential for both normal people and patients (Akyuz and Kirbag 2010; Daba et al., 2008).Oyster mushrooms are cultivated widely as their temperature requirement of 20-30C prevails in most of the areas. From 1997 to 2010, Pleurotus spp. production increased from 876 t to 6,288 t (618%). China was responsible for most of the production increase and accounted for over 85% of the world’s total output in 2010. (Royse, 2014 )

 Edible mushrooms are grown on waste materials from farms, plantations agro factories recycled to produce value-added mushrooms. Currently, millions of tons of agricultural wastes are discarded, burned and neglected thus reducing environmental pollution (Bhawnaand Thomas, 2003). These agro wastes can be used alone or in combination to create mushroom growing substrates (Chang, 1991 in an inexpensive way.

Grevillaerobusta (Proteaceae), commonly known as “Silky Oak” is native to Australia (Richieet al., 1965; Cannon et al., 1973). G. robusta is popular among farmers in Kenya, due its fast growth, ability to tolerate heavy pollarding and pruning of branches and because it mixes well with other crops (Tyndall,1996;Muchinet al. 2002). The species has important uses including construction material, fuel wood, shade, fodder soil erosion control, and soil fertility improvement (FAO, 2001;Holdinget al., 2005). So far, alkyl resorcinols, macrocyclic phenols and cinnamic acid derivatives have been reported to be constituents of this plant. In Ethiopia, Grevillaerobustais a dominant tree planted as a road side shade throughout the country, in different institutions including the capital city, Addis Ababa. The use of this tree for the various purposes is not yet promoted for the scale it required.  This article discusses about the usability of Grevillaerobusta dry leaves together with the different proportion of cotton seed waste for the cultivation of oyster (Pleurotusostreatus) mushroom.

 

 

MATERIALS AND METHODS

 

Organism and culture conditions

 

The fungal strain, Pleurotusostreatus(Oyster mushroom) was obtained from Mycology Laboratory, Department of Biology, Addis Ababa University, Ethiopia. The pure culture of Pleurotusostreatuswas transferred on to Potato Dextrose Agar (PDA) on the Petri dishes  and incubated at 25C. The growth of the culture and presence of contamination were visually inspected at three days interval.

 

Grain Spawn production

 

The spawn (mushroom seed) of Pleurotusostreatuswas produced on yellow colored sorghum grain, wheat bran and calcium sulfate (gypsum) in the ratio of 88:10:2 respectively (Dawit, 1998). The required amount of sorghum grain was weighed and soaked over night in sufficient amount of water. The grains were washed and drained to remove the dead and floating seeds with excess of water. After removing the excess water from the grain, the required amount of wheat bran and gypsum (CaSO4 2H20) were added and transferred to 1000 ml glass bottles (75% level) leaving a head space over the grain and autoclaved at 121°C temperature for 45 minutes. After cooling, each bottle was inoculated with 20 agar blocks (1 cm × 1 cm) of 15day old mushroom culture from the Petri dish and incubated for 21 days at 28 ± 2C until the substrate were fully colonized and the mycelia invasion and contamination were inspected at five days interval.

 

Treatments

 

Nine treatments (T1–T9) comprising different proportions of Grevillaerobusta dry leaves, waste paper and cotton seed waste (600 g) with 1% lime stone (Calcium Carbonate) on dry weight basis were used as shown in Table 1.

 

 

Table1: The composition of different treatments used in this study

Treatments

Grevillae

dry leaves (g)

Cotton seed

waste (g)

Waste 

Paper(g)

Total (g)

T1

450

50

100

600

T2

400

100

100

600

T3

350

150

100

600

T4

300

200

100

600

T5

250

250

100

600

T6

200

300

100

600

T7

150

350

100

600

T8

100

400

100

600

T9

50

450

100

600

 

 

Preparation of the substrate

 

The Grevillae dry leaves were socked in sufficient amount of water over night. On the next day excess water was squeezed from the leaves. The waste paper was cut into small pieces approximately (3–5 cm), weighed and soaked in sufficient amount of water immediately before use. Excess water present in the substrates was drained thoroughly and mixed with required amount of wheat bran and one percentof calcium carbonate and filled in sterilizable yellow color polyethylene bags (Kurtupestal). The substrates were autoclaved at 15Psi pressure at 121ºC temperatures for 1hour. After sterilization the substrates were transferred to transparent polyethylene cultivation bags for easy supervision of the growth of the mycelia and presence of contamination. Each substrate (600 g) with 70% moisture was mixed with 10% spawn (dry weight/wet weight basis) and the inoculated polythene bags were then tightly tied with string made from polyester/cotton cloth. Pin holes were made through the bags (1/100 cm2) for drainage and aeration. It was kept in a spawn running room at room temperature.After primordial formation, the bags were transferred to mushroom house under normal environmental conditions and relative humidity (the room maintained at 85–90%),irrigated using tap water every morning and evening with optimum ventilation. The mushrooms were manually harvested at maturity which was indicated by up ward curving of the edges of the cap. Biological efficiency was calculated and defined as the ratio of weight (g) of fresh mushrooms harvested to dry weight (g) of the substrate (Chang et al., 1981)

 

 

Data analysis

 

The data were analyzed by comparing the mean weights and percent biological efficiency through one way ANOVA. The data groups were analyzed using Statistical Package for Social Sciences (SPSS) for windows version 20.  Treatment mean were compared using LSD.

 

 

RESULTS AND DISCUSSION

 

Mycelia colonization of the substrate

 

The production bags that received different substrates mixture showed significantly different mycelia colonization rate (p≤ 0.05).Treatment 9,8 and 7 showed relatively short period (10 days) for mycelia colonization than  treatments 5 and 6 which took 19 days (Table.2). Sharma et al., (2016) reportedcomplete of mycelia colonization from 22.40-26.00 days.The time required for first primordial formation in the different treatment showed no significant variation(p≥ 0.05) (Table 2).  In this study, the pin head formation after complete mycelia colonization took longer time than the duration reported by Sharma et al., (2016)that is 6 days in fastest and 8 days in slowest treatments.Kumari and Achal (2008) stated that colonization of the substrate was completed within 20 days of inoculation. Quimioet al. (1999) reported that good harvest of P. ostreatuswas 3-4 weeks after incubation. Shah et al. (2004) reported that primordial formation of P. ostreatusappears 27-34 days of inoculation which is longer than the results of this study.The total days required for complete production cycle of the oyster mushroom using Grevillae dry leaves as major substrate showed significant (p≤ 0.05) differences. Zinabuet al., (2015) reported that sugarcane bagasse supported the fast mycelial growth during cultivation ofPleurotus species. In general, fastest mycelia run and shortest time taken for complete colonization of the substrate, the different mixes of the substrates satisfy the required amount of assimilable nutrients for the mycelia growth.

Treatment 8 relatively took shorter total days for complete production while treatment 3, 5, 6, took more days for completing the  production cycle.

 

Table2: Duration of the different phases of growth of the mushroom

Treatment

Complete mycelia

colonization

First primordial formation

after colonization

Completion of production(days)

T1

16b

7 a

110

T2

16b

7 a

104

T3

16b

7 a

111

T4

16b

7 a

110

T5

19c

8 a

111

T6

19c

8 a

111

T7

10a

6 a

110

T8

10a

6 a

100

T9

10a

6a

106

 

 

Yield (fresh weight) of mushroom in different harvesting cycle

 

The yields of mushroom biomass during the first harvest showed significant (p≤ 0.05)variationin in  the different treatments. During this harvest treatment 9 with less amount of Grevillae dry leaves gave the highest yield(570g) and treatment 1 gave the lowest yield (275g)(Table 3). All the remaining treatments showed fresh weight of mushroom between the highest and lowest.During the second cycle harvest, treatments 7,8 and 9 showed high rate of biomass,346,310and 320g respectively. The biomass harvested declined progressively from the third to the fifth harvest invariably with all treatments. However, relatively good harvest was observed with treatments 4,8 and 9.This observation is  in line with reports of Asefa  and Lakew (2016) and  Sharmaet al., (2016) that  highest biomass was found  in the first flush than the rest of the harvest.

 

 

Table 3: Yield (fresh weight) of mushroom in different harvesting cycle

Treatments

1st flush 

2nd Flush

3rd flush

4thflush

5thflush

T1

275i

140h

73h

50f

-

T2

350g

207 f

190a

84e

88b

T3

300h

238e

109g

94d

73c

T4

415e

278d

174b

125b

105a

T5

440d

213 f

117f

105 c

60d

T6

400f

210f

151d

130b

88b

T7

500 b

346a

140e

108 c

75c

T8

450c

310c

165c

145a

75c

T9

570a

320b

178b

107c

73c

 

 

Relatively shorter pinning to maturation were observed on treatments 8,7, and 9. While more days form pinning to maturation were taken for the treatments of 5 and 6 (Table 4).  In all rest of harvesting cycle (2-5) there were no significant ((p≥ 0.05) variation on the days taken from pinning to maturation. According to Mekonnen and Semira, (2014) oyster mushroom grown on cotton hull substrates took 4 and 7 days for first and second harvesting time respectively than the saw dust substrate. The combination of substrate with less amount cotton seed waste showed shortest mean periods from pinning to maturation from first to fourth flushing in the order 10, 9, 8, and 7days respectively as compared to other treatments. Similarly short pinning to maturation period of 4 days was reported by Gumeet al.(2013) for oyster mushroom grown on sawdust and coffee waste.

The average number of fruits of the treatments bags that received different substrate mix showed significant differences (p≤ 0.05).Treatment 4 and 7 showed the highest number (average) of fruits  130 each while treatment 1 gave the lower number of fruits  70 (Fig 1).  And the rest of the treatments gave number of fruits between the highest and the lowest. The result recorded in this study was in line with Sharma et al. (2016)) who indicated that  the number of fruit bodies produced in the first harvest was higher than the subsequent flushes.

 

Table 4:Duration from pinning to maturation(days)during harvesting cycle

Treatments

1st flush 

2nd  Flush

3rd  flush

4th flush

5th flush

T1

8

7

7

7

-

T2

8

7

7

7

6

T3

8

7

7

7

6

T4

8

7

7

7

6

T5

9

8

7

7

6

T6

9

8

7

7

6

T7

7

6

6

6

6

T8

7

6

6

6

6

T9

7

6

6

6

6

 

 

 

The average number of aborts of the treatments bags that received different substrate mix showed significant variations (p≤ 0.05).The treatment 4 and  9 showed lower number of aborts 60 each, while   treatment 8showed highest number of aborts 91. And the rest of the treatments showed number of aborts between the highest and lowest(Fig 1).

Average number of bunches in the various  treatment mix showed significantvariations (p≤ 0.05) The treatments 9 (15), 6 ( 14) and 4 (14) showed relatively higher number of bunches  while treatment 1 (7) had fewer number of bunches(Fig 1).The average cap diameter with treatment 6 was greater than treatment 1 (Fig 2).This observation was in line with, the observation of Sharma etal., 2016)., who indicated that, the size of fruiting bodies was higher in case ofrice straw  than in all other substrates.

Stipe length: the average stipe length of the treatments bags that received different substrate mix showed significantly different (p≤ 0.05).treatment 3, 8, 5 and 9 gave (relatively shorter stipe length which were ranged  from 2.6-2.8 cm while, treatment 1, 2, and 7 gave relatively longer stripe length3.5 cm each. And treatment 4 and 6showed intermediate stipe length between the longer and shorter length 3.0 each (Fig 2).

 

 

Fig 1: Number of aborts, bunches and fruits

 

 

Fig 2:Cap diameter and stipe length of the different treatments

 

 

Total biomass and biological efficiency

 

The highest total biomass was recorded on treatment 9( 1246.5g fresh weight /600g of the dry substrate)while the lowest biomass was recorded on treatment 1, (538 g of fresh mushroom /600 g of dry substrate)(Fig 3). The highest biological efficiency was also recorded from treatment 9, (208)and  the lowest from treatment 1( 89.5%)(Fig 4). Zinabuet al., (2015) reported  high values of biological efficiency for Pleurotusostreatus grown on Sugarcane bagasse  (70.5%) and Pleurotusostreatus grown on waste paper  alone (68.9%) followed by Pleurotusflorida grown on sugarcane bagasse (67.6%).Patra and Pani (1995) reported biological efficiency (50-75%) of Pleurotus species grown on agro-industrial residues, namely; corncobs, various grasses and reed stems, vine shoots, cottonseed hulls and sugarcane bagasse. In another report Sharma et al., (2016) observed thehighest yield (381.85gm) and biological efficiency (95.46%)  using  rice straw , wheat straw, paper and sugarcane bagasse while lowest yield 247.87g and biological efficiency 61.96 % was obtained from sawdust.Royse  et al(2004)stated that, yields for cottonseed hull/wheat straw substrate were three fold higher than treatments containing switch grass. The present study results are in agreement with their observation that as the proportion of cotton seed waste increased in the substratum the total biomass yield and biological efficiency also increased.

 

 

Fig 3:Totalbiomass of the different treatments

 

 

Fig 4: The Biological efficiency of the different treatments

 

 

Fig 5: A) GrevillaerobustaB) Grevillae dry leaves, C) cotton seed D) waste paper: E)oyster mushroom culture on dextrose agar medium, F) spawn prepared on yellow colored sorghum grain ready to use, G) Fully colonized substrate in the production bag, H) Primordial formed on the production bag, I) Mature fruit bodies of oyster mushroom

 

 

CONCLUSION

 

Mushroom production technology has been considered as appropriate technology  for satisfying  nutritional needs and medicinal requirements and for the sustainability of the environment through recycling of organic waste which otherwise negatively affect the environment. Large bodies of information have been available on the usability of different agricultural and agro processing by products for the cultivation of mushrooms. But very few reports were available on processing dry foliage for the production of mushroom. In this study the usability of Grevillaerobusta dry leaves  along with cotton seed waste , and waste paper were used as substratum at different proportions. The total yield of the mushroom biomass, as well as the biological efficiency showed significant difference between the treatments. However, it is evident that Grevillaerobustae dry leaves can be utilized as a mushroom substratum along with other agricultural by-products for production of mushrooms. 

 

 

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Cite this Article: Asefa K (2018). Evaluation of Grevillae Robusta Dry Leaves Supplemented with Cotton Seed Waste for the Cultivation of Oyster mushroom (Pleurotusostreatus). Greener Journal of Agricultural Sciences, 8(6): 110-118, http://doi.org/10.15580/GJAS.2018.6.051518071.