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21 
J. mt. area res., Vol. 1, 2016 

   

       Journal of Mountain Area Research 
    

 

 
EVALUATION OF GIN WASTE AS A GROWING SUBSTRATE, ENRICHED WITH 

DIFFERENT VOLUME PERCENTAGE OF THE WHEAT BRAN FOR CULTIVATION 

OF OYSTER MUSHROOM (PLEUROTUS OSTREATUS) 
 

M. M. Akhtar1, *, M. M. Jahangir2, M. Zeeshan1, M. Abdullah1 

 

1. World Wide Fund for Nature Pakistan (WWF-Pakistan) 

2. Institute of Horticultural Sciences, University of Agriculture Faisalabad, Pakistan 

 

 

 

ABSTRACT 

 Pleurotusostreatus mushroom was cultivated on cotton gin waste amended with wheat bran in 

order to judge its growth potential. Two substrates (cotton gin waste and wheat bran) were 

employed alone and with different combinations. Experiment consisted of four treatments T0 (100 % 

cotton gin waste), T1(97% cotton gin waste + 3% wheat bran), T2(94% cotton gin waste + 6% wheat 

bran) and T3(91% cotton gin waste + 9% wheat bran). Data about time needed for commencement 

of spawn run, time needed for completion of mycelial growth, time needed for initiation of pinheads, 

time needed for harvesting of 1st, 2nd and 3rd flush, fresh weight of 1st, 2nd and 3rd flush harvested, 

total yield, pH of mushroom, total soluble solids of mushroom, acidity and ascorbic acid contents, 

reducing sugars, non-reducing sugars and total sugars of mushroom, total nitrogen, phosphorus and 

potassium contents of mushroom was recorded. T0 (100 % cotton gin waste) performed better as 

compared to other treatments. 

 

KEYWORDS: Cotton gin waste, wheat bran, biochemical, yield, mushroom growth 

 

 

*Corresponding author: (Email: makhtar@wwf.org.pk) 

 

1. INTRODUCTION 

Oyster Mushroom Pleurotusostreatus is a 

famous mushroom and is an important part of 

daily diet of the people in various countries of 

the world like; China, Korea and Japan. This 

mushroom grows wildly in forests of hilly areas 

and flourishes better at temperature range of 

22-28°C and atmospheric humidity more than 

85%. In Pakistan, Oyster Mushroom grows 

naturally on trunk of trees and stumps in 

Northern Areas, Azad Jamu and Kashmir [1]. 

Oyster Mushroom is popular enriched source 

of proteins, vitamins such as B, D, E, and K (fats 

(0.5-3.5%) and various health beneficial [2]. In 

Pakistan production of Oyster Mushroom is 

getting popularity due to its high economic 

returns. Globally farmers cultivate Oyster 

Mushroom on different agricultural waste 

materials such as wheat straw, rice straw, 

sorghum straw, rice husk, alang-alang grass, 

artichoke waste, azolla, banana leaves, 

banana pseudostems, bean pods, bean 

straw, cactus, agave and yucca, cardamon 

pulp, cinnamon leaves, peels of citrus fruit, 

coconut husks, coconut coir and coconut 

pith, coffee parchment, coffee sawdust corn 

fiber, crushed corncobs, hammer milled corn 

Vol. 1, 2016 

http://journal.kiu.edu.pk/index.php/JMAR 

Full length article 

mailto:makhtar@wwf.org.pk


 

 

 

Akhter et al., J. mt. area res. 01 (2016) 21-27 

22 J. mt. area res., Vol. 1, 2016 

 

cob, corn stapes, corncobs, corn leaves, corn 

stover, corn stalks, lemon grass leaves, oat 

straw, ragi straw, sesame stems, sugarcane 

bagasse sunflower husks, sunflower peels, 

chopped heads and stipes of sunflower and  

tea leaves etc. [3]. 

 

The Cotton Gin Waste is normally 

composed of leaves, sticks, soil particles, burs, 

other plant materials, cotton lint and mote 

etc. One of the biggest problems faced by 

the cotton ginning industry in Pakistan and rest 

of the world is Cotton Gin Waste 

Management. It has been estimated that 

ginning one bale (227 kg) of spindle harvested 

seed cotton lint approximately contributes 

between 37 and 147 kg of waste [4]. As 

reported by the Pakistan Cotton Ginners 

Association (PCGA), total cotton production 

in Pakistan in 2012-13 stood at 12.915 million 

bales, approximately 12.81% or 1.898 million 

bales less than previous year. Considering that 

on the average annually about 12 million 

bales are ginned in Pakistan, the amount of 

cotton gin waste produced in Pakistan could 

be approximately close to 3.5 billion pounds 

per year.  

 

Disposal of such huge amount of cotton 

gin waste is an alarming issue as it may cause 

serious threats to the environment. At the 

moment, global cotton industry is trying to 

reduce their cotton gin waste by alternative 

options for handling this as a by-product, 

which has potential as a multiuse product.  

Use of cotton gin waste as a growing 

media for Oyster Mushroom cultivation could 

be a viable option in this regard. As growing 

of Oyster Mushroom on cotton gin waste can 

be helpful in reducing environmental pollution 

caused by cotton gin waste. The objective of 

this study was to investigate the possibility of 

Oyster Mushroom production on cotton waste 

and wheat bran and their different 

combinations. 

 

2. METHODOLOGY 

 

Present research work was accomplished 

in mushroom farm of a village in Basti Misson 

Bahawalpur, in a project entitled “Farmers’ 

capacity building for mushroom growing using 

cotton gin waste as growing substrate” under 

WWF-Pakistan’s SPRING project “Sustainable 

Cotton Production in Pakistan’s Cotton 

Ginning SMEs” funded by European Union. 

While all the biochemical analysis was 

performed at Institute of Horticultural 

Sciences, University of Agriculture, Faisalabad 

during 2013-2014 for evaluation of growth and 

yield response of Oyster Mushroom specie viz. 

Pleurotusostreatus by using cotton gin waste 

mixed with wheat bran at different ratios. 

 

Cotton waste was used as substrate in this 

research work. Cotton gin waste was soaked 

in water. Two percent lime was mixed in 

cotton gin waste to maintain its pH. After 

soaking, the substrate was piled up and 

covered with polythene sheet. Cotton gin 

waste was allowed to ferment for 4 days. 

Cotton gin waste was then spread on floor for 

evaporation of excess water. Latter on wheat 

bran was added @ 3, 6 and 9% respectively. 

Substrate was filled in polypropylene bags of 

size (7×9 inch) and bags mouths were loosely 

tied with rubber bands. The bags were 

pasteurized by local method in an ordinary 

drum for two hours. When bags were 

pasteurized, they took one day for cooling. 

After cooling the bags were inoculated with 

spawn at the rate of 10gm per bag. During 



 

 
 

Akhter et al., J. mt. area res. 01 (2016) 21-27 

23 
J. mt. area res., Vol. 1, 2016 

spawn running the temperature in growth 

room was controlled between 22-26°C for 

spawn running. The required humidity was 

maintained between 70-80% by sprinkling 

water on the floor several times a day. 

 

After completion of spawn running the 

temperature of growing room was maintained 

between 16-250°C. Fructification or fruit body 

was started as soon as the substrate was fully 

impregnated with mycelial growth. The 

humidity of the growing room was maintained 

between 80-90% by sprinkling water on floor 

and moisture requirements of the bags was 

accomplished by sprinkling water on them 

thrice a day using sprinkler. 

 

Experiment was conducted under 

complete randomized design and every 

treatment was replicated five times. 

T0= Cotton waste (100%) control 

T1=Cotton waste (97%) + Wheat bran (3%) 

T2= Cotton waste (94%) + Wheat bran (6%) 

T3= Cotton waste (91%) + Wheat bran (9%) 

 

Data regarding different parameters like 

time taken for commencement of spawn run 

(days), time taken for completion of mycelial 

growth (days), time taken for initiation of 

pinhead formation (days), Number of days for 

completion of 1st  flush (days), Number of 

days for completion of  2nd  flush (days), 

Number of days for completion of 3rd  flush 

(days), fresh weight of 1st flush (g), fresh 

weight of 2nd flush (g), fresh weight of 3rd  

flush (g), Total  yield (g), pH, Total soluble 

solids (°Brix), acidity (%) [5], Ascorbic acid 

(mg/100ml) [6], reducing sugars, non-

reducing sugars, total sugars [5], notrogen 

contents of mushrooms, phosphorus contents 

of mushrooms and potassium contents of 

mushrooms [7] was commemorated. 

2.1 Statistical Analysis  

In this experiment completely randomized 

design (CRD) was used. The data collected 

sequentially was examined statistically using 

LSD test at 5% probability level [8]. 

 

3. RESULTS AND DISCUSSION 

Data regarding effect of cotton gin 

waste, wheat bran and their different 

amendments on growth of Oyster Mushroom 

(Pleurotusostreatus) is shown in Table 1. The 

minimum time for commencement of spawn 

run was taken by T0(1.4 days) while T3 took 

maximum time (2.4 days) for commencement 

of spawn run followed by T2 and T1 

respectively. Likewise, Time taken for 

completion of mycelial growth was 

commemorated in days. Treatments showed 

significant results regarding completion of 

mycelial growth. The Treatment T0 performed 

well and took minimum time (22 days), 

followed by T1(30.2 days), T2 (42.4 days) and 

T3 (47 days) respectively for completion of 

mycelial growth. Time taken for initiation of 

pinhead formation was also commemorated 

in days. T0 took minimum time (26.8 days) for 

initiation of pinheads followed by T1(34.8 

days),T2(47 days)and T3(52 days) respectively 

as shown in Table 1. An experiment done by 

Khan et al. [9] support our work as they 

reported better performance of Pleurotus 

species on cotton waste as compared to 

different other mushroom growth media. The 

fluctuation in above cited parameters may be 

ascribed to fluctuation in lignocellulosic and 

chemical composition of growing substrates 

[10]. 

 



 

 

 

Akhter et al., J. mt. area res. 01 (2016) 21-27 

24 J. mt. area res., Vol. 1, 2016 

 

Table 1. Effect of substrate on time taken for 

completion of mycelia growth of Oyster Mushroom 

(Pleurotusostreatus). 

Treatments Time taken for 

commencement  

of spawn run 

(days) 

Time taken 

for 

completion 

of mycelial 

growth 

(days) 

Time taken for 

initiation of 

pin head 

formation(da

ys) 

T0 1.4000 C 22.000D 26.800 D 

T1 1.8000 BC 30.200C 34.800 C 

T2 2.000 AB 42.400B 47.000 B 

T3 2.4000 A 47.000A 52.000 A 

LSD Value 0.5996 2.5963 2.4540 

 

Figures not sharing the same letters differ significantly 

at P = 0.05. 

Data concerning time taken for 

completion of 1stflush, Time taken for 

completion of 2ndflush and Time taken for 

completion of 3rdflush is shown in Table2. T0 

(31.2 days) took minimum time for completion 

of 1stflush followed by T1(42.2 days),T2(43.2 

days), and T3(55.8 days) respectively. 

Likewise, T0 (39.2 days) took minimum time for 

completion of 2ndflush followed by T1(49.2 

days),T2(51.4 days)and T3(62.8 days) 

respectively. Likewise, T0 (46.8 days) took 

minimum time for completion of 3rdflush 

followed by T1(56.2 days),T2(57.2 days)and 

T3(69.8 days) respectively. Similar behavior of 

mushroom harvesting pattern was reported by 

Dundar and Yildiz [11]. 

 

 

 

 

 

 

Table 2. Effect of Substrate on number of days for 

completion of flush of Oyster Mushroom 

(Pleurotusostreatus). 

 

Treatments Number of 

days for 

completion 

of 1st  flush 

Number of 

days for 

completion of 

2nd  flush 

Number of 

days for 

completion of 

3rd  flush 

T0 31.200 C 39.200  C   46.800C 

T1 42.200 B 49.200 B 56.200B 

T2 43.200 B 51.400 B 57.200B 

T3 55.800 A 62.800 A 69.800A 

LSD Value 3.8974 

 

3.8743 4.0667 

Figures not sharing the same letters differ significantly 

at P = 0.05. 

 

Data regarding Fresh weight of 1stflush 

(g), Fresh weight of 2ndflush (g), Fresh weight 

of 3rdflush (g) and Total yield (g) is shown in 

Table 3. In case of fresh weight of 1stflush 

harvested T0 yielded best (76.2 g) followed by 

T1(63.4 g),T2(50 g) andT3(42.2 g) respectively. 

While, in case of fresh weight of 2ndflush 

harvested T0 yielded best (48.6 g) followed by 

T1(39.75 g),T2(31.20 g)and T3(22.8 g) 

respectively. Similarly, in case of fresh weight 

of 3rdflush harvested T0 yielded best (35.99 g) 

followed by T1(28.31 g),T2(20.95 g)and 

T3(15.76 g) respectively. On the other hand, in 

case of total yield T0 performed best (159.8 g) 

followed by T1(131.39 g),T2(107.86 g)and 

T3(80.76 g) respectively. With the passage of 

time gradual decrease in mushroom yield was 

noticed from 1st flush to 3rd flush which might 

be ascribed to gradual reduction in nitrogen 

contents of substrates [12]. 

 

 

 



 

 
 

Akhter et al., J. mt. area res. 01 (2016) 21-27 

25 
J. mt. area res., Vol. 1, 2016 

Table 3. Effect of substrate on yield parameters of 

Oyster Mushroom (Pleurotusostreatus). 

Treatments Fresh weight 

of 1st flush 

(g) 

Fresh 

weight of 

2nd  

flush(g) 

Fresh 

weight of 

3rd 

flush(g) 

Total 

yield (g) 

T0 76.200 A 48.600 A 35.998  A 159.80 A 

T1 63.400 A 39.756 AB 28.31B 131.39 B 

T2 50.000 B 31.204 BC 20.952C 107.86 C 

T3 42.200 B 22.800 C 15.76D 80.76 D 

LSD Value 13.046 10.975 4.0811 16.991 

Figures not sharing the same letters differ significantly 

at P = 0.05. 

 

Data regarding pH, Total soluble solids 

(0Brix), Acidity of mushroom (%) and Ascorbic 

acid contents of mushroom (mg/100ml) is 

shown in Table 4. Highest pH value was 

observed in case of T3 (7.96) followed by 

T2(7.84),T1 (7.7)and T0(7.54) respectively. In 

case of total soluble solids of mushroom 

maximum total soluble solids were observed in 

case of T0 (3.6) followed byT1 (3.3) and T2 

(3.19) and T3(3.02) respectively. In case of 

acidity of mushroom Maximum acidity of 

mushroom was observed in case of T0 (0.042) 

followed by T1(0.036), T2(0.034) and T3(0.032) 

respectively. In case of ascorbic acid 

contents of mushroom, maximum ascorbic 

acid contents were observed in case of T0 

(10.61 mg/100ml) followed by T1(8.2 

mg/100ml), T2 (7.4 mg/100ml) and T3 (7.0 

mg/100ml) respectively. Mycelial growth is 

maximum at high pH while it is minimum at low 

pH [13]. 

 

Table 4. pH of substrate their amendments on 

biochemical attributes of Oyster Mushroom 

(Pleurotusostreatus). 

Treatments PH Total 

soluble 

solids TSS 

(°Brix), 

Acidity (%) Ascorbic 

acid 

(mg/100ml) 

T0 7.542 B 3.6000 A 0.0429A 10.614 A 

T1 7.7000 

AB 

3.3000 A 0.0365AB 8.200 A 

T2 7.8400 

AB 

3.1980 A 0.0346AB 7.400 A 

T3 7.9600 

A 

3.0200 A 0.0322A 7.000 A 

LSD Value 0.3082 0.6475 8.394 3.6749 

Figures not sharing the same letters differ significantly 

at P = 0.05. 

 

Data regarding reducing sugars contents 

of mushroom (%), non-reducing sugars of 

mushroom (%) and total sugar contents of 

mushroom (%) is shown in Table 5. In case of 

reducing sugars contents of mushroom (%), 

maximum reducing sugars contents were 

observed in case of T0 (3.44%) followed by 

T1(3.35%), T2(3.21%) and T3(3.15%) 

respectively. Similarly, in case of non-reducing 

sugars contents of mushroom (%), maximum 

non reducing sugars contents were observed 

in case of T0 (6.99%) followed by T1(6.18%), 

T2(6.03%) and T3(5.96%) respectively. similarly, 

in case of total sugars contents of mushroom 

(%), maximum total sugars contents were 

observed in case of T1 (10.51%) followed by 

T0(10.49%), T2(9.97%) and T3(9.34%) 



 

 

 

Akhter et al., J. mt. area res. 01 (2016) 21-27 

26 J. mt. area res., Vol. 1, 2016 

 

respectively. Maximum Non reducing sugars, 

reducing sugars in mushroom were observed 

which grow on substrate having cotton wastes 

only [13]. 

 

Table 5. Effect of substrate and their amendments on 

reducing sugars, non-reducing sugars and total 

sugars of Oyster Mushroom (Pleurotusostreatus). 

 

Treatments Reducing 

sugars(%) 

Non reducing sugars 

(%) 

 

Total 

sugars (%) 

T0 3.4440 A 6.9980 A 10.492  A 

T1 3.3580 AB 6.1840 B 10.518  A 

T2 3.2180 AB 6.0340 B 9.9780 AB 

T3 3.1520 B 5.9680 B 9.3460  B 

LSD Value 0.2298 

 

0.6131 0.8380 

Figures not sharing the same letters differ significantly 

at P = 0.05. 

 

Data about nitrogen contents of 

mushroom (%), phosphorus contents of 

mushroom (%) and potassium contents of 

mushroom (%) is shown in Table 6. In case of 

nitrogen contents of mushroom (%), maximum 

nitrogen contents were observed in case of T0 

(0.71 %) followed by T1(0.69%), T2(0.60%) and 

T3(0.50%) respectively. Similarly, in case of 

phosphorus contents of mushroom (%), 

maximum phosphorus contents were 

observed in case of T0 (0.45 %) followed by 

T1(0.43%), T2(0.31%) and T3(0.24%) 

respectively. Likewise, in case of potassium 

contents of mushroom maximum potassium 

contents were observed in case of T0 (0.76 %) 

followed by T1(0.70%), T2 (0.57 %) and 

T3(0.56%) respectively. It is reported that 

Phosphorus, potassium and sodium are vital 

minerals of mushrooms [14]. 

 

Table 6. Effect of substrate and their amendments on 

Nitrogen, Phosphorus and Potassium contents of 

Oyster Mushroom (Pleurotusostreatus). 

Treatments Mushroom 

Nitrogen 

contents (%) 

Mushroom 

Phosphorus 

contents (%) 

Mushroom 

Potassium 

contents 

(%) 

T0 0.7140 A 0.4540 A 0.7660 A 

T1 0.6900 A 0.4360 A 0.7080 A 

T2 0.6000 B 0.3140 B 0.5780 B 

T3 0.5000 C 0.2460 C 0.5600 B 

LSD Value 0.0864 0.0425 0.1076 

Figures not sharing the same letters differ significantly 

at P = 0.05. 

 

4. CONCLUSION 

 It can be concluded from the present 

research that T0, which is 100% cotton gin 

waste used as growing substrate, performed 

best as compared to other Treatment. So, 

cotton gin waste can be successfully used for 

cultivation of high quality Oyster Mushroom 

(Pleurotusostreatus). 

 

References 

[1] Z. Sher, F. Hussain and L. Badshah. Micro-

mineral contents in eight forage shrubs at three 

phenologicaltages in a Pakistan’s rangeland. 

African Journal of Plant Science, 5 (2011) 557-

564 

[2]  M. Bozena, S. Katarzyna, L. Mciej, O. 

Wlodzenriers, Z. Magdalena, R. Jack. Edible 



 

 
 

Akhter et al., J. mt. area res. 01 (2016) 21-27 

27 
J. mt. area res., Vol. 1, 2016 

Mushrooms in Prophylaxis and Treatment of 

Human Diseases. J. MIR. 4 (2013)170-183  

[3]  M. S. Sangwan and L. C. Saini. Cultivation of 

Pleurotussajorcaju (Fr.) Singer on agro-industrial 

wastes. Mushroom Res. 4 (1995) 33–34. 

[4] J. A. Thomasson, A Review of Cotton Gin Trash 

Disposal and Utilization. Beltwide Cotton 

Conferences: (1990)689-705. 

[5] W. Hortwitz, Official and Tentative Methods of 

Analysis, 9(1960) 320–41. Association of the 

Official Agriculture Chemist. Washington, D.C. 

[6] J. A. Ruck, Chemical Methods for Analysis of 

Fruits and Vegetables. Research Station 

Summerland, Canada Department of 

Agriculture, (1969) Publication No:1154 

[7] D. H. Chapman and F. Parker,  Determination 

of NPK. In: Methods of analysis for soils, plants 

and water. 1st Ed. California University, 

Agriculture Division, USA. (1961) 150-179. 

[8] R.G.D. Steel, J. H. Torrie and D.A. Dickey. 1997. 

Principles and Procedures of Statistics: A 

Biometrical Approach. 3rd ed. McGraw Hill 

Book Co., New York. 

[9] N. A. Khan, M. I. Haq and M. A. Khan. Growth 

comparison of local and exotic strains of oyster 

mushrooms on different agricultural wastes. 

Pak. J. Phytopathol. 21 (2009) 139-143. 

[10]  E. A. Adebay and D. Martinez- Carrera. Oyster 

Mushrooms (pleurotus)are useful for utilizing 

lignocellulosic biomass. African Journal of 

Biotechnology, 14 (2015) 52-67 

[11]  A. Dundar and A. Yildiz. A comparative study 

on Pleurotusostreatus (jacq.) P. kumm. 

cultivated on different agricultural 

lignocellulosic wastes. Turk. J. Biol., 33 (2009) 

171-179. 

 

 

 

 

[12] M. M. Jahangir, G. Samin, N. A. Khan, A. 

Rehman, Z. Aslam, M. Amjad, K. Ziaf, R.W.K. 

Qadri, I. Khan and M. Atiq. Performance of 

Oyster Mushroom (Pleurotus) grown on cotton 

waste and sorghum straw based growing 

substrate. Pak. J. Phytopathol. 27 (2015) 77-81. 

[13]  H. Sardar, M. A. Ali, C. M. Ayyub, and R. Ahmad. 

Effect of different culture media, temperature 

and pH level on the growth of wild and exotic 

pleurotus species. Pak. J. Phytophathol. 25 

(2015) 139-145. 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

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