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Am. J. Environ. Clim. 1(1), 2022

The American Journal of  Environment and Climate (AJEC) is a blind peer-reviewed journal, that aims to publish 
the article(s) immediately after submission of  the corrected version by the author. The journal publishes articles that 
include those containing substantially supported theories, innovative works, substantial experimental results, and/or 
containing useful and constructive discussions or reviews standardized to regional or international acceptance. The 
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internet immediately upon receiving the final versions.

Frequency: Three issues per year 
Area of  publication: Environmental Science & Technology, Environment Pollution, Policy  on Environment and 
Climate, Climate Change, Sustainable Development.

Editorial Team

Dr. Muhammad Farhad Howladar
Department of  Petroleum and Mining Engineering
Shahjalal University of  Science and Technology Sylhet, Bangladesh
Professor Anand V. Shivapur
Department of  Civil Engineering Visvesvaraya Technological University
Belagavi, Karnataka state, India
Dr. Sofia Binte Ehsan
Institute of  Climate Change, Universiti Kebangsaan Malaysia, Malaysia
Dr. Velichko Dimitri Vladimirobitch
Russian State Geological Prospecting University, Russia
Dr. K. J. Sreekanth
Research Scientist, Energy Efficiency Technologies Program (EET)
Energy and Building Research Center (EBRC)
Kuwait Institute for Scientific Research (KISR) Safat, Kuwait
Dr. Mohammad Tofayal Ahmed
Department of  Petroleum and Mining Engineering,
Jessore University of  Science and Technology, Bangladesh
Dr. Asif  Rayhan
National University of  Malaysia, Malaysia
Dr. Md. Mahfuzul Islam
Institute for Environment and Development
Univeriti Kebangsaan Malaysia, Malaysia
Ar. Sajal Chowdhury
Department of  Architecture
Chittagong University of  Engineering & Technology (CUET), Bangladesh
Dr. Parnuwat Usapein
Acting Director of  Rattanakosin College for Sustainable Energy and Environment
Rajamangala University of  Technology Rattanakosin (RMUTR), Thailand
Dr. Vikrant Katekar
Research Scholar, Indian Institute of  Technology, Mumbai, India
Dr. Sheikh Tawhidul Islam
Professor & Director, Institute of  Remote Sensing
Jahangirnagar University, Bangladesh

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American Journal of  
Environment and Climate (AJEC)

Growth and Yield Performance of  Oyster Mushroom Cultivated in Combined Cassava 
Peels, Coconut Residue and Coffee Waste Substrates

Irish B. Elsisura1*, Mary Amor G. Figueroa2

Volume 1 Issue 1, Year 2022
ISSN: 2832-403X (Online)

DOI: https://doi.org/10.54536/ajec.v1i1.206
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Article Information ABSTRACT

Received: March 22, 2022

Accepted: April 1, 2022

Published: April 8, 2022

The oyster mushroom (Pleurotus ostreatus) is an edible mushroom that belongs to the class 
of  Basidiomycetes. It has reached sufficient market maturity because of  its flavor, shelf-life 
durability, and protein and fiber content. Besides their nutritional, medicinal, and economic 
value, they may help the country’s agricultural waste management, bridge environmental 
issues, and contribute to climate change resolution advancements. A study on different 
varieties of  agricultural substrates derived from waste materials such as cassava peels, 
coconut residue, and coffee waste was investigated and compared to sawdust, the common 
substrate for oyster mushrooms. The effects of  different substrates on the morphological 
characteristics of  P. ostreatus, percent contamination, and yield parameters were recorded 
and analyzed using the Analysis of  Variance in Completely Randomized Design, and 
their significant results were compared using Tukey’s HSD. Results showed that different 
substrate mixtures did not significantly influence the morphological characteristics of  P. 
ostreatus. Moreover, sawdust, the common substrate for oyster mushrooms, showed the 
lowest percent contamination as compared to other substrate mixtures. Contaminants found 
in cassava substrates include Trichoderma spp., Aspergillus spp., Fusarium spp., Neurospora spp., 
and Penicillium spp. 80% of  cassava peels combined with 10% coconut residue and 10% 
coffee waste significantly increased the number of  fruiting bodies and produced the heaviest 
fresh weights of  oyster mushrooms. Stipe length and pileus diameter were also significantly 
influenced by this substrate mixture, which is comparable to the common substrate. 
However, further research on the varying proportions of  these substrate mixtures on the 
performance of  oyster mushrooms is recommended.

Keywords
Cassava peel, Coconut Residue, 
Coffee waste, Oyster mushroom, 
Growing media

1 Student Researcher of  Surigao State College of  Technology, Mainit Campus, Magpayang,  Mainit, Surigao del Norte, Philippines
2 Faculty Adviser of  Surigao State College of  Technology, Mainit Campus, Magpayang, Mainit, Surigao del Norte, Philippines
* Corresponding author’s e-mail: ielsisura.tc@ssct.edu.ph

INTRODUCTION
Oyster mushrooms have reached sufficient market 
maturity because of  their delightful flavor, durability 
in shelf  life, extraordinary protein and fiber content, 
and nutritional and medicinal features. These kinds 
of  mushrooms can establish and degrade a variety 
of  lignocellulosic substrates and other wastes, which 
are produced primarily through the activities of  the 
agricultural, forest, and food-processing industries.
Because of  the more favorable climate conditions in 
Southeast Asian countries such as the Philippines, growing 
one’s food poses few to no challenges. The country’s 
main crops are rice, corn, coconut, sugarcane, bananas, 
pineapple, coffee, mangoes, tobacco, and abaca. Secondary 
crops include peanuts, cassava, sweet potatoes, garlic, 
onion, cabbage, eggplant, lime, lemon, rubber, and cotton. 
These are all export-quality products that the country is 
most proud of. But with the assortment of  waste materials 
it produces, it piles it up to the extent of  harming the 
environment itself, which supplies all of  it, ironically. 
But the ballooning effect of  our population reflects the 
agricultural and waste management of  our country. 
Today, only a part of  this waste is used, mainly as 
provisions for goats and other pastured animals. In many 
regions where cassava is cultivated and processed, volumes 
of  cassava peels are dumped along the roads, especially 
in wet seasons, and can generate offensive odors and 
unsanitary conditions. A valorization of  cassava waste 

would not only resolve this environmental predicament 
but also contribute to the local economy.
The coconut residue generated after milk extraction is a food 
waste by-product, but only a fraction of  it is used as fertilizer 
or cow feed. As a result of  these activities, the by-products 
can contribute to the additional trash in the community.
By-products from coffee beans are useful substrates for 
the cultivation of  P. ostreatus in coffee-producing countries 
like the Philippines because this species is fast-growing, 
substrates are easily accessible, and the mushrooms (or 
fruiting bodies) are a beneficial source of  nutrition and 
revenue.
The researcher recognized the fact that there was not 
enough identified data that involved the previously 
indicated substrate combinations in developing P. 
ostreatus, although individual studies have shown 
that these substrates are effective in growing oyster 
mushrooms as previously mentioned. P. ostreatus is 
uncommon in our area, and the researcher was strongly 
convinced that the need to discover and innovate ways 
to cultivate it was significant. It was anticipated in this 
research to compare and contrast the yield performance 
of  oyster mushroom (P. ostreatus) cultivation in a rural 
setting using different agricultural waste materials such as 
cassava wastes, coconut residue, and used coffee grounds, 
along with a constant percentage of  supplementations 
from rice bran, limestone, and molasses and sawdust 
as common substrate. The researcher compared five 

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substrate mixtures based on the yield performance in 
the production of  oyster mushrooms. Subsequently, the 
study determined which of  these substrates produced the 
most viable yield.
Through this study, Philippine households and farmers, 
particularly those in Mainit, Surigao del Norte, can have 
the opportunity and means to learn more about cultivating 
edible mushrooms such as oyster mushroom. Making 
these concepts a reality on a micro-to-commercial scale 
can suit the people’s nutritional demands, not only as a 
supplement but also with profitable future engagements. 
It can also provide an opportunity for everyone with 
the resources to address and contribute to the country’s 
current understanding of  agricultural waste management 
not only ecologically but also economically.

LITERATURE REVIEW
For numbers of  years, oyster mushroom (P. ostreatus) 
has displayed as one of  the most cultivated mushrooms 
in the world, largely in Brazil (Lee et al., 2002; Sánchez, 
2010; Royse, 2013). Pleurotus species’ nutritional value, 
comprises high protein of  25-50 %, nine essential amino 
acids, and very low 2-5 % fat content, which makes it 
fitting as a diet food for health-conscious individuals. The 
sugar content is reasonable and ranges 17-47%, including 
minerals such as calcium, potassium, sodium with vitamins 
such as niacin, riboflavin, vitamin B1, B5, B6, C, and D 
(Caglarirmak, 2007; Syed et al., 2009).  In addition, oyster 
mushrooms are good source of  extraordinary stamina 
and vigor, and they are utilized in the manufacturing 
of  many continental recipes. They also have medical 
properties such as anti-cancer, anti-cholesterol, and anti-
tumorous. Mushrooms can help with diabetes, ulcers, 
and lung illness (Quimio, 1976). Mushroom protein is a 
combination of  animal and vegetable protein (Kurtzman, 
1976). Mushrooms are also potent in Niacin, Pantothenic 
acid, and Biotin (Subramanian, 1986). The good thing is, 
we can easily grow it on agricultural and industrial waste.  
More than half  of  the overall yield from the land is wasted 
as straws, leaves, stalks, roots, and so on (Zadrazil, 1978). 
These pollutants can be repurposed into food, and the 
environment may be less polluted as a result (Hayes, 1978). 
The population of  the Philippines is climbing to 
110,623,413 (as of  Sunday, March 21, 2021; based on 
web). With this increase in population is the increase 
in the demand for food and agricultural products. As 
a result, it had been estimated that only about 50% of  
the city household’s solid wastes are pulled by garbage 
collectors, while 38% of  the households incinerate their 
garbage, and a further 12% of  it is dumped in vacant 
lots (Holmer et al., 1999). The bigger portions of  the 
solid waste that are being dumped at the city landfill site 
are from private households (54%) and commerce and 
institutions (28%). About 40 to 50% of  the city wastes 
are biodegradable. However, if  organic waste is properly 
processed to form compost, it can be put to good use 
in urban agriculture and horticulture as a fertilizer and 
soil improver. Farmers and gardeners can benefit from 

it diligently. The use of  organic waste provides a lasting 
increase in the waste control situation in cities. However, 
this assumes that urban waste management is integrated 
consistently with urban horticulture and agriculture 
(Guanzon & Holmer, 2015).
In Thailand, they can easily grow oyster mushrooms (P. 
ostreatus) in local conditions if  the appropriate specifications 
of  food and moisture for growth are available (Shah et al., 
2004). Pleurotus ostreatus demands few environmental controls, 
and their fruiting bodies are not often attacked by diseases 
and pests, and they can be cultivated simply and cheaply. All 
this makes P. ostreatus cultivation an excellent alternative for 
the production of  mushrooms when compared to other 
mushrooms (Sánchez, 2009). 
Cassava (Manihot esculenta) belongs to the higher 
classification Manihot; and in order Malpighialesis, which 
is a dominant food crop for approximately 700 million 
people, especially in African countries. A substantial 
quantity of  waste is usually produced throughout the 
processing, mainly consisting of  tuber peels (Sonenberg 
et al., 2015). Previous reports on the utilization of  cassava 
waste for the cultivation of  P. ostreatus and P. pulmonarius 
have shown that yields were reasonable (Adebayo et al., 
2009; Onuoha et al., 2009; Obodai et al., 2014). Cassava 
by-products as substrates can give yields of  up to 100 
percent BE of  oyster mushrooms which compare well to 
traditional substrates such as sawdust (Frimpong-Manso 
et al., 2011). Cassava-based composites thus have the 
potential to be economically profitable for the production 
of  oyster mushrooms (Sonnenberg et al., 2015).
Coconut palm (Cocos nucifera L.) is a tropical plant that 
belongs to the Kingdom Plantae, division Magnoliophyta, 
class Liliopsida, order Arecales, and family Arecaceae. 
The adult de-husked coconut generates 50% wet meat or 
core, 33% shell, and 17% water. Raw coconut meat has 
a chemical composition that contains about 3.33 percent 
protein, 33.49 % total fat, 15.23 % starch, 3.23 % total 
fiber, and a range of  minerals and vitamins. The meat 
of  the coconut kernel is used to make coconut milk in 
mostly Asian counties like the Philippines. Fresh coconut 
kernel is finely shredded and hand-squeezed or expeller 
pressed to produce coconut milk. As a result, a significant 
volume of  coconut waste by-product after squeezing out 
milk is razed to the ground as waste (Sopit, 2007). 
Coffee grounds have been investigated as a potential 
remnant for mushroom growth. Coffee grounds’ 
chemical and structural properties enable for the reuse 
of  lignin (23.90 g/100g dry product), nitrogen (12.79 
g/100g dry product), and dietary fiber (60.40 g/100g 
dry product). The storage capacity of  water is influenced 
by the particulate matter produced by coffee pressing 
(Ropciuc et al.,2016). Oyster mushroom cultivation on 
SCG for human consumption is totally possible, and there 
is significant potential for this extensive form of  waste 
to be utilized and the caffeine content reduced. (Cabrera, 
2018). Lime (CaCO3) is a key component in mushroom 
development; commercial mushroom production is 
dependent on correct substrate pH adjustment. (Khan 

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et al., 2013). Furthermore, molasses has been shown 
to promote the growth of  several bacteria. Molasses 
contains sugar, nitrogen, and other nutrients that aid in 
cell development. As a result, it produced good results in 
the production of  oyster mushrooms. (Erkel, 2009). Rice 

bran is clearly the most widely used and widely available 
organic component in substrates for the production of  a 
wide range of  edible mushrooms (Peng et al., 2000)

MATERIALS AND METHODS
Table 1. Research material, equipment, and raw materials used.
Materials Equipment Raw materials
Metal strainer Improvised mushroom bag sterilizer Cassava peels
Cheesecloth Shovel Coconut residue
6x12 inches Polypropylene cellophane Shredder Coffee waste
Empty bottle Pressurized sprayer Sawdust
PVP pipe (2.0 cm thick) Triple burner gas tank Limestone
Cotton wool Water drum Molasses
Rubber bands Rice bran
Soap Water
Bleach
Small-eyed net
Basin
Tarpaulin
Black cloth
Pail
Spoon
70% Alcohol 
Denatured alcohol
Denatured alcohol lamp
Sacks
LED light bulb

The following materials, equipment, and raw materials 
were used in the study (Table 1).
Acquisition of  Pleurotus ostreatus Grain Spawn and 
Substrate’s Raw Materials
The oyster mushroom spawn was procured from a 
commercial laboratory located in Taguibo, Butuan City.
Different raw materials needed for preparing substrates 
for the study were gathered from the local area in Mainit, 
Surigao del Norte. Specifically, cassava wastes and coconut 
residue were from the farmers and neighborhood within 
Mainit, Surigao del Norte. The coffee-pressed grounds 
were collected from the nearest coffee shop within 
Surigao City, Surigao del Norte.
Preparation of  Mushroom Substrates  
Cassava wastes. Fresh cassava wastes were carefully 
cleaned with water, packed into nylon sacks and were 
submerged in water for three days to mimic natural 
fermentation. After three days, the fermented cassava 
wastes were drained for a day, and sun-dried for two days 
using a net or a table. The sun-dried cassava wastes were 
shredded using a shredder. When the desired texture and 
sizes were attained, it was then packed in a clean sack and 
stored in a dry place.
Coconut residue. Finely shredded fresh coconut meats 
were soaked in warm water. The soaked shredded coconut 
meats were hand-squeezed and pressed through a metal 

strainer, and cheesecloth. In order to get lighter coconut 
milk, the process was repeated once or twice. After the 
milk extraction, the coconut leftovers were sun-dried for 
at least two days. The dried coconut residues were stored 
in a cool and dry place.  
Coffee grounds. Coffee-spent grounds that were gathered 
from a coffeehouse were dumped in hot water to eliminate 
chemicals that might limit fungal development and 
trigger contamination. To eliminate the extra moisture, 
cheesecloth was used to softly pressed it to allow any 
leftover moisture to travel through. The coffee waste was 
left to cool off  in a room temperature for about a day or 
two. It was placed in a dry container and stored in a cool 
and airy corner.
Building and Maintenance of  Mushroom House
The mushroom house was four meters from floor to top 
part, and with an area of  four-square meters. It only has 
one door, no window, and was made of  bamboo matting 
with a nipa roof.
The mushroom house was cleaned thoroughly in its 
entirety by using pressurized water hose with soap, and 
bleach to ensure the removal of  almost every unwanted 
bacterium or foreign biological substance that could alter 
the growing process and was regularly sanitized.
Preparation and Bagging of  Mushroom Substrates
Recently prepared substrates were packaged in a 6x12 

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inch polypropylene cellophane bags containing 500 grams 
each based on the different substrate mixtures (Table 2). 
The bottom ends of  the bags were folded to allow them 
to stand on their own. As additional compost was added 
to the ultimate weight required, an empty bottle was used 
to compress it. To act as a bottle neck, a 2.0 cm thick and 
2.5 cm long PVC pipe was put at the neck of  each bag. 
A piece of  cotton wool was inserted into the bags’ necks 
then covered with paper, and was secured with a rubber 
band. Then, it was labelled carefully and accordingly 

before proceeding to the next step.
Sterilization of  Bagged Substrates  
Bagged substrates were sterilized for six hours using the 
improvised mushroom bag sterilizer, fired with a triple 
burner stove. The sterilized bags were then removed 
from the improvised mushroom bag sterilizer to allow 
the second batch to be accommodated.
After sterilization, the substrates were allowed to totally 
cool-off  for six to ten hours before spawn inoculation.    

Table 2. Experimental treatments.
Treatment 
Numbers

Oyster Mushroom Substrate Mixtures Total
Major Substrate Ingredients (78%) Supplemental Ingredients
Sawdust
(Control)

Cassava 
peels

Coconut 
residue

Coffee 
waste

Rice bran Molasses Lime-
stone 

1 100%  - - - 20% 1% 1% 100%
2 - 100% -  - 20% 1% 1% 100%
3 - 80% 10% 10% 20% 1% 1% 100%

4 - 70% 20% 10% 20% 1% 1% 100%
5 - 50% 40% 10% 20% 1% 1% 100%

Grain Spawn Inoculation to Sterile Bagged Substrates
The oyster mushroom grain spawn was inoculated to the 
sterile bagged substrates in a biosafety cabinet. The grain 
spawn was loosened first before inoculation by shaking it. 
Bags were opened carefully, grain spawn were aseptically 
inoculated to the substrates at one tablespoon per bag, 
and then was covered again for incubation. 
Incubation of  Inoculated Fruiting Bags
The inoculated fruiting bags were arranged properly 
by treatments and replication, then they were placed in 
a well-ventilated dark area, covered in black cloth, and 
layered by tarpaulin or any plastic material to prohibit 
the direct light to our fruiting bags, until fully colonized. 
Fruiting bags were regularly monitored for colonization 
period.
Hanging of  Fully-colonized Fruiting Bags
After colonization, the bags were sorted and hung using 
a rope and a wire as a lock to the thread. To depict each 
replication, fifteen mushroom fruiting bags were arranged 
in a row of  every treatment piled at one foot apart. These 
were then regularly monitored until the appearance of  
pinheads’ formation.
Management of  Fruiting Bags
Spraying water at the back portion of  the fruiting bags 
were done twice a day, at nine o’clock in the morning 
and three o’clock in the afternoon. The fruiting bags were 
enclosed in a net to avoid insect and pests’ infestations. 
The floor was completely covered with a damp cloth to 
keep the mushroom house’s humidity level stable. The 
time pinheads were observed, the cotton and paper cap 
of  the fruiting bags were removed to allow the fruiting 
body to develop. During the stage of  fruiting body 
formation, the room was illuminated with diffused 
light. Bags were checked for contamination on a daily 
basis. To prevent the spread of  contaminants, defective 
bags were disposed or eradicated as soon as they were 

observed. Contaminations were identified based on their 
morphological characteristics and were compared to 
published and identified common contaminants of  oyster 
mushrooms.
Harvesting of  Oyster Mushroom Fruits
Harvesting was done when the cap of  the oyster 
mushroom reached its maturity with its maximum 
diameter and in convex shape, by hand picking. Fruiting 
bodies were twisted and pulled carefully from the base of  
the stem to avoid contamination.  
Data Gathered
During the course of  the study, the following parameters 
were recorded.
1. Morphological characteristics. This parameter includes 
period of  colonization or spawn run, pinhead    
formation, fruiting body formation and full maturity of  
oyster mushroom.

a) Days from Spawning to Complete Spawn Run 
(S-CSR). This refers to the number of  days the   
substrates were fully colonized by the fungus from the 
day of  inoculation.

b) Days from Spawning to Pinhead Formation (S-PF). 
This pertains to the number of  days the fungus  
produces pinheads’ formation from the day of  inoculation

c) Days from Pinhead Formation to fruiting body 
formation (PF- FBF). This refers to the number   
of  days the fruiting body was observed after the pinhead 
formation, and 

d) Days from Fruiting Body Formation to Full Maturity 
(FBF- FM). This represents the ideal number   
of  days to harvest after fruiting body formation. 
2. Percentage Contamination. It refers to the percentage 
of  discarded fruiting bags due to contamination   
in every treatment. 
This was computed using the formula: 

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3. Harvest Data. This parameter includes number of  
fruiting bodies per treatment, average mycelia height,  
average mycelia diameter, and weight of  fresh fruiting 
bodies of  oyster mushroom on different substrate 
mixtures.

a) Number of  fruiting bodies. Immediately every after 
harvest, fruiting body was counted by determining the 
caps of  the fruits.

b) Average stipe height. It was gathered by measuring 
the tip of  the stem or the stipe of  the oyster    
mushroom to the base of  the cap. A ruler was used and 
was expressed in centimeters.

c) Average pileus diameter. The pileus or the cap of  the 
oyster mushroom was measured across using a ruler and 
was expressed in centimeters.

d) Weight of fresh fruiting bodies of oyster mushroom. It 
was gathered every after harvest. Electronic   
balance was used to gather the data and was expressed in grams. 
Statistical Analysis
The study was analyzed using Analysis of  Variance 
(ANOVA) in Completely Randomized Design and the 
differences among treatment means were compared using 
Tukey’s Honest Significant Difference (THSD) as Post-
Hoc analysis.

RESULTS AND DISCUSSION
Morphological Characteristics of  P. ostreatus 
Grown in Different Substrate Mixtures
Days from Spawning to Complete Spawn Run (S-CSR). 
The colonization period of  oyster mushroom using 
different substrate mixture is presented in Table 3. 
Results of  the Analysis of  Variance (ANOVA) showed 
no significant differences. Results of  the study showed 
that 100% cassava peel has the shortest colonization 
period (33.33 days from inoculation), which was followed 
by 80% cassava peels + 10% coconut residue + 10% 
coffee waste with 34.67 days of  colonization period, and 
70% cassava peels + 20% coconut residue + 10% coffee 
waste with 35 days of  inoculation. The longest period of  
colonization was recorded in substrate mixtures of  100% 
sawdust (Control) and 50% cassava peels + 40% coconut 
residue + 10% coffee waste with both 35.33 days. These 
results suggest that different substrate mixtures did not 
influence the period of  colonization of  P. ostreatus.

Days from Spawning to Pinhead Formation (S-PF). The 
pinhead’s formation period of  oyster mushroom using 
different substrate mixtures is also presented in Table 3. 
Same results of  the ANOVA were recorded showing no 
significant differences among treatment means. Shortest 
pinhead formation was recorded in substrate mixtures 
with 100% cassava peels with 37.33 days. Eighty percent 
cassava peels + 10% coconut residue + 10% coffee 
waste and 70% cassava peels + 20% coconut residue 
+10% coffee waste have shorter period of  pinhead 
formation with both took 38.00 days from inoculation. 
These were followed by the control (100% sawdust) 
with 38.67 days of  pinhead formation. Substrate mixture 
with 50% cassava peels + 40% coconut residue + 10% 
coffee waste registered the longest number of  days of  
pinhead formation with 40.00 days from inoculation. 
However, general results of  this parameter suggest that 
these substrate mixtures did not significantly influence 
the pinhead formation of  P. ostreatus.
Days from Pinhead Formation to Fruiting Body 
Formation (PF- FBF). The period of  fruiting body 
formation of  oyster mushroom using different substrate 
mixtures is presented in Table 3 as well. ANOVA results 
showed no significant differences among treatment 
means. Data showed that whatever substrate mixtures 
used, there is no differences in period of  fruiting body 
formation which only took three days from pinhead 
formation in all substrate mixtures.
Days from Fruiting Body Formation to Full Maturity 
(FBF- FM). The time taken from fruiting body formation 
to pin maturation is also presented in Table 3. Results of  
the ANOVA revealed no significant differences among 
treatment means. Results of  the study showed that all 
substrate mixtures used had no significant influenced 
on the period of  full maturation of  P. ostreatus which 
only took three days from fruiting body formation in all 
substrate mixtures. 
Generally, results imply that different substrate mixtures 
used in the experiment do not influence the colonization 
period, pinhead formation, fruiting body formation, and 
full maturation period of  P. ostreatus. These findings could 
be attributed to the cellulose and lignin properties of  
both cassava peels and sawdust (Frimpong-Manso et al., 
2011) (Horisawa et al., 1999). The results also confirmed 
Stanley & Nyenke’s (2011) findings that cassava 
stimulated luxuriant mycelial growth rate and extension 

Table 3. Morphological characteristics of  oyster mushroom in different treatments.
Substrate Mixtures S-CSR 

1,ns
S-PHF 2,ns PHF-FBF 

3,ns
FBF-MP 
4,ns

100% Sawdust (Control) 35.33 38.67 3 3
100% cassava peels 33.33 37.33 3 3

80% cassava peels + 10% coconut residue + 10% coffee waste 34.67 38.00 3 3
70% cassava peels + 20% coconut residue + 10% coffee waste 35.00 38.00 3 3
50% cassava peels + 40% coconut residue + 10% coffee waste 35.33 40.00 3 3
CV (%) = 2.68 2.52 0.00  0.00

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can be comparable to that of  a commonly used substrate 
such as sawdust. 
ns=not significant
1 =S-CSR (Days from Spawning to Complete Spawn 
Run)
2 =S-PHF (Days from Spawning to Pinhead Formation)
3 =PHF-FBF (Days from Pinhead Formation to Fruiting 
Body Formation)
4 =FBF-MP (Days from Fruiting Body Formation to Full 
Maturity)
Days from Spawning to Pinhead Formation (S-PF). The 
pinhead’s formation period of  oyster mushroom using 
different substrate mixtures is also presented in Table 3. 
Same results of  the ANOVA were recorded showing no 
significant differences among treatment means. Shortest 
pinhead formation was recorded in substrate mixtures 
with 100% cassava peels with 37.33 days. Eighty percent 
cassava peels + 10% coconut residue + 10% coffee 
waste and 70% cassava peels + 20% coconut residue 
+10% coffee waste have shorter period of  pinhead 
formation with both took 38.00 days from inoculation. 
These were followed by the control (100% sawdust) 
with 38.67 days of  pinhead formation. Substrate mixture 
with 50% cassava peels + 40% coconut residue + 10% 
coffee waste registered the longest number of  days of  
pinhead formation with 40.00 days from inoculation. 
However, general results of  this parameter suggest that 
these substrate mixtures did not significantly influence 
the pinhead formation of  P. ostreatus.
Days from Pinhead Formation to Fruiting Body 
Formation (PF- FBF). The period of  fruiting body 
formation of  oyster mushroom using different substrate 
mixtures is presented in Table 3 as well. ANOVA results 
showed no significant differences among treatment 
means. Data showed that whatever substrate mixtures 
used, there is no differences in period of  fruiting body 
formation which only took three days from pinhead 
formation in all substrate mixtures.
Days from Fruiting Body Formation to Full Maturity 
(FBF- FM). The time taken from fruiting body formation 
to pin maturation is also presented in Table 3. Results of  
the ANOVA revealed no significant differences among 
treatment means. Results of  the study showed that all 
substrate mixtures used had no significant influenced 
on the period of  full maturation of  P. ostreatus which 

only took three days from fruiting body formation in all 
substrate mixtures. 
Generally, results imply that different substrate mixtures 
used in the experiment do not influence the colonization 
period, pinhead formation, fruiting body formation, and 
full maturation period of  P. ostreatus. These findings could 
be attributed to the cellulose and lignin properties of  
both cassava peels and sawdust (Frimpong-Manso et al., 
2011) (Horisawa et al., 1999). The results also confirmed 
Stanley & Nyenke’s (2011) findings that cassava 
stimulated luxuriant mycelial growth rate and extension 
can be comparable to that of  a commonly used substrate 
such as sawdust.
Percent Contamination of  Oyster Mushroom Bags 
Prepared With Different Substrate Mixtures
Percent of  contamination. The percent contamination of  
oyster mushroom bags prepared with different substrate 
mixtures is shown in Table 4. Results of  the ANOVA 
revealed significant differences among treatment means 
(p > 0.01). The least percentage of  contamination was 
recorded in the control bags with 100% sawdust (11.11%), 
followed by the substrate mixtures of  80% cassava peels 
+ 10% coconut residue + 10% coffee waste and 50% 
cassava peels + 40% coconut residue + 10% coffee 
waste comparable percent contamination of  37.78% and 
35.56%, respectively. Higher contamination was observed 
in substrate with 100% cassava peels with 71.11% 
contaminated bags. While the highest contamination was 
recorded in substrate mixtures with 70% cassava peels + 
20% coconut residue + 10% coffee waste with 84.44% 
contamination.  
Common contaminants found in the contaminated 
fruiting bags were presented in Table 5. The fungal 
contaminants observed were as follows: Trichoderma spp., 
Aspergillus spp., Fusarium spp., Neurospora spp., and Penicillium 
spp. These results suggest that 100% sawdust as substrate 
for P. ostreatus had the lowest percent of  contamination. 
Hence, it can be the ideal substrate for oyster mushroom 
to lower contaminant population. Moreover, cassava 
mixed with coconut residue and coffee waste can 
enhance contaminant population. These may be due to 
the fungal contaminants usually associated with higher 
percentage of  cassava-based substrate like Aspergillus spp. 
(Obadina, 2006), and coconut-based substrate Fusarium 
spp. (Manimekalai, 2010). Number of  

Table 4. Percent of  contaminated oyster mushroom bags in different substrate mixtures.
Substrate Mixtures Replication Total Mean**

I II III
100% Sawdust (Control) 6.67 13.33 13.33 33.33 11.11a

100% cassava peels 66.67 73.33 73.33 213.33 71.11c

80% cassava peels + 10% coconut residue + 10% coffee waste 33.33 40.00 40.00 113.33 37.78b

70% cassava peels + 20% coconut residue + 10% coffee waste 86.67 86.67 80.00 253.33 84.44d

50% cassava peels + 40% coconut residue + 10% coffee waste 33.33 33.33 40.00 106.67 35.56b

CV (%) = 8.02; **=significant at 1% level, THSD.

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Table 5. Contaminants found in contaminated fruiting bags

Substrate Mixtures Image of  
Contaminations

Morphological 
Characteristics

Fungal Contaminants

100% 
Sawdust (Control)

Green-mold.
Color yellow to green, 
gradually creeping into the 
black substrate
(Druzhinina et al., 2011)

Trichoderma spp.

100% 
cassava peels

Produced white to yellowish 
or crusty on the substrate, 
foul-rotten smell.
(Lopez - Arevalo et al. 
1996))

Aspergillus spp

80% cassava peels + 10% 
coconut residue + 10% 
coffee waste

Fusarium mold, can appear 
pale or brightly colored, 
with a cottony surface.
(Manimekalai, 2010)

Fusarium spp.

70% cassava peels + 20% 
coconut residue + 10% 
coffee waste

Produced bright neon 
orange to reddish substrate.
(Suada et al.2015)

Neurospora spp.

50% cassava peels + 40% 
coconut residue + 10% 
coffee waste

Produced initially white and 
become blue-green, gray-
green, olive-gray, yellow or 
pinkish with time substrates.
(Lopez - Arevalo et al. 
1996))

Penicillium spp.

Fruiting Bodies of  Oyster Mushroom 
Grown in Different Substrate Mixtures
Number of  fruiting bodies produced in different substrate 
mixtures is presented in Table 6. Results of  ANOVA 
revealed significant effects of  these substrates to fruiting 
bodies of  oyster mushroom on its first, second, and 
total flushes. Effects of  the number of  fruiting bodies is 
shown in Fig. 1.
First Flush. Results of  the study showed that fruiting 
bodies (43.15) were significantly higher in substrates 
with 70% cassava peel + 20% coconut residue + 10% 
coffee waste which was comparably higher as compared 
to substrate with 80% cassava peel + 10% coconut 
residue + 10% coffee waste with 37.25 fruiting bodies 
formed, and was followed by the substrate with 100% 
cassava peels with 34.30 fruiting bodies. Lower number 
of  fruiting body formation was observed in substrate 
with 50% cassava peel + 40% coconut residue + 10% 
coffee waste with 30.70 fruits formed. Lowest number 
of  fruiting bodies on the other hand, was recorded in 
control (100% sawdust) with only 23.21 fruits formed. 
Second Flush. Results showed that 50% cassava peel + 
40% coconut residue + 10% coffee waste had the highest 
number of  fruiting bodies with 50.09 fruits formed, 
which is comparably higher as compared to substrate 
with 80% cassava peel + 10% coconut residue + 10% 

coffee waste and 100% cassava peels with 46.56 and 35.00 
fruits formed. Seventy percent of  cassava peels + 20% 
coconut residue + 10% coffee waste and the control 
(100% sawdust) had the lowest number of  fruiting bodies 
(19.75 and 21.49 fruits, respectively).
Total Flush. Results showed that 80% cassava peel 
+ 10% coconut residue + 10% coffee waste and 50% 
cassava peel + 40% coconut residue + 10% coffee waste 
had the highest number of  fruiting bodies with 83.81 
and 80.78 fruits, respectively. These were followed by 
the substrate with 100% cassava peels and 70% cassava 
peel + 20% coconut residue + 10% coffee waste having 
corresponding fruit formations of  69.30 and 62.90. 
Lowest number of  fruiting bodies was recorded in the 
control (100% sawdust) with only 44. 70 fruits.
Generally, the above results suggest that the substrate 
mixtures containing 80% cassava peel + 10% coconut 
residue + 10% coffee waste and 50% cassava peel + 
40% coconut residue + 10% coffee waste can increase 
the production of  fruiting bodies of  oyster mushroom 
for about 36.08 - 39.11. These corresponds to the 
outcomes of  the study of  Sonnenberg et al. (2015) that 
demonstrated the potential of  cassava peel as a substrate 
ingredient for the production of  oyster mushrooms, 
with a yield comparable to that of  sawdust. These results 
were further explained by Youri (2003), who showed that 

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sawdust and cassava peel are lignocellulosic material which 
consist of  three main components, namely: cellulose, 
hemicellulose, and lignin that serve as a suitable substrate 
for mushrooms as it degrades lignocellulosic substrates 
through lignocellulosic enzyme production and utilize the 
degraded products to produce their fruiting bodies, and 
were also backed up by the studies of  Sa´nchez (2009), 
and Grim & Wosten (2018). Furthermore, Ginterova and 
Janotkova (1998) discovered that plant oil with a high 
total fat content, such as that found in coconut residue, 

stimulates the formation of  biomass in P. ostreatus. As a 
result, adding the appropriate amount of  coconut residue 
can be used as supplemental substrate, increasing the 
fruiting bodies and yield of  oyster mushroom (Sopit, 
2007). Besides, Ropciuc et al. (2016), also observed that 
mushrooms grown on coffee waste had a respectable 
percentage of  biological efficiency. As a result, combining 
these substrates can contribute to achieving the highest 
yield in weight parameters of  oyster mushroom.

Table 6. Yield of  oyster mushroom in different substrate mixtures.
Substrate
Mixture

No. of  Fruiting Body
(Per flush)

Stipe 
Height 
(cm) **
   

Pileus 
Diameter 
(cm) *

Fresh Weight
(g. per flush)

1st * 2nd ** Total** 1st** 2nd**
100% Sawdust (Control) 23.21d 21.49c 44.70c 19.45a 21.15a 37.99bc 38.10b

100% cassava peels 34.30b 35.00b 69.30b 11.35bc 13.90ab 50.00ab 51.00a

80% cassava peels + 10% coconut 
residue + 10% coffee waste

37.25ab 46.56ab 83.81a 12.17b 14.34ab 63.63a 53.50a

70% cassava peels + 20% coconut 
residue + 10% coffee waste

43.15a 19.75c 62.90b 11.00c 13.40ab 52.10ab 32.25b

50% cassava peels + 40% coconut 
residue + 10% coffee waste

30.70c 50.09a 80.78a 9.44d 10.78b 32.18c 34.25b

CV (%) =  19.84 14.34 12.31 2.91 21.88 13.97 7.69

*=significant at 5% level, THSD; **=significant at 1% level, THSD.

Figure 1. Number of  fruiting bodies of  oyster 
mushroom grown in different substrate mixtures.

Stipe Height (cm) and Pileus Diameter (cm) of  
Oyster Mushroom Grown in Different Substrate 
Mixtures
Stipe height (cm) and pileus diameter (cm) of  oyster 
mushroom grown in different substrate mixtures is 

also presented in Table 6. Results of  ANOVA revealed 
significant differences on both parameters.
Stipe height (cm).  Results showed that the control (100% 
sawdust) has the longest stipe height with 19.45 cm. This 
was followed by the substrate with 80% cassava peel + 
10% coconut residue + 10% coffee waste with 12.17 cm. 
height which is comparable with 100% cassava peels with 
11.35 cm. stipe height. Shorter stipe height (11.00 cm.) 
was observed in 70% cassava peel + 20% coconut residue 
+ 10% coffee waste which as followed by the substrate 
with 50% cassava peel + 40% coconut residue + 10% 
coffee waste with 9.44 cm.
Pileus Diameter (cm). Results of  the study showed control 
(100% sawdust) still showed the widest pileus with 21.15 
cm. Comparably, wider pileus were observed in substrate 
mixtures such as 100% cassava peels, 80% cassava peel 
+ 10% coconut residue + 10% coffee waste and 70% 
cassava peel + 20% coconut residue + 10% cof-fee waste 
with corresponding pileus diameter of  13.90 cm., 14. 
34 cm., and 13.40 cm. Smallest pileus was recorded in 
substrate containing 50% cassava peel + 40% coconut 
residue + 10% coffee waste with 10.78 cm. wide.
The above results imply that 100% sawdust has the 
longest stipe and widest pileus with 19.45cm. and 
21.15 cm., respectively. Comparable pileus size was 
also observed in substrates such as 100% cassava peels, 
80% cassava peel + 10% coconut residue + 10% coffee 
waste, and 70% cassava peel + 20% coconut residue + 
10% coffee waste. These results inclined to the study of  
Sanjel et al. (2021), that discovered that the size of  the 

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mushroom is dependent on substrates, and that poor 
in cellulose, hemicelluloses, and lignin materials can 
constitute physical barriers that are difficult to be broken 
down without the presence of  lignin-degrading enzymes. 
In addition, major ecological factors that affects the stipe 
height, and pileus diameter in oyster mushroom includes 
the compactibility or the compressibility of  material used 
as substrate. Because of  the particle size, it increases 
the void ratio as moisture plays an important role in 
bonding because of  sawdust and cassava peel’s cohesive 
or lignin characteristics (Azhar et al., 2015). It has been 
supported by several studies that substrate type such as 
its lignin characteristic contributes significant differences 
in stipe height, and pileus diameter of  oyster mushroom 
(Besufekad et al., 2020; Nkwonta, 2013; Tsegaye & Tefera, 
2017; Onyeka et al., 2018; Dubey et al., 2019).

Figure 2. Pileus diameter (left), and stipe height (right) 
of  oyster mushrooms harvested from different substrate  
mixtures.

Fresh Weights (g) of  Oyster Mushroom as 
Influenced by Different Substrate Mixtures
Fresh weights (g) of  oyster mushroom grown in different 
substrate mixtures is presented in Table 6. Results of  the 
ANOVA revealed significant effects in the fresh weight 
of  oyster mushrooms in first and second flushes. 
First Flush. Heaviest fresh weight (63.63 g) was recorded 
in oyster mushroom grown from substrate with 80% 
cassava peel + 10% coconut residue + 10% coffee 
waste. Comparable effects were observed in substrates 
containing 100% cassava peels, and 70% cassava peel 
+ 20% coconut residue + 10% coffee waste with 50.00 
grams and 52.10 grams, respectively. Lighter fresh weights 
on the other hand, were recorded in substrates with 100% 
sawdust (37.99 g.) and 50% cassava peel + 40% coconut 
residue + 10% coffee waste (32.18 g.). 

Second Flush. Heaviest fresh weight of  oyster mushroom 
on the second flush were observed in 100% cassava 
peels and 80% cassava peel + 10% coconut residue + 
10% coffee waste with weights of  51.00 grams and 53.50 
grams, respectively. Lighter fresh weights were recorded 
in the other substrate mixtures such as 100% sawdust, 
70% cassava peel + 20% coconut residue + 10% coffee 
waste, and 50% cassava peel + 40% coconut residue + 
10% coffee waste with weights that ranged from 32.25 
grams – 38.10 grams.
Based on these results, it suggests that 80% cassava peel 
+ 10% coconut residue + 10% coffee waste can produce 
a heavy fresh weights of  oyster mushroom. Moreover, 
comparable results can be obtained using 100% cassava 
peels as substrate. These results confirmed the study of  
Sonnenberg et al. (2015) that cassava peels showed to 
be a potential ingredient in mushroom substrate for it 
produced more than 100% BE which is comparable to 
that the usual sawdust substrate. Moreover, several reports 
showed that the adding of  coconut residue (Sopit, 2007), 
and coffee waste (Ropciuc et al., 2016) increases yield and 
gained a respectable percentage of  biological efficiency 
in oyster mushrooms. Therefore, combination of  these 
substrates can contribute to gaining the significant yield 
in weight parameters of  oyster mushroom. 

CONCLUS ION
Based on the above results, it can be concluded that various 
substrate mixtures do not significantly influence the oyster 
mushroom’s colonization, pinhead formation, fruiting 
body formation, and the period of  maturation. Among 
other substrate mixtures, the common substrate (100% 
sawdust) can produce the least number of  contaminated 
fruiting bags, as well as the widest mushroom pileus and 
longest stipe. Moreover, the substrate mixture with 80% 
cassava peels + 10% coconut residue + 10% coffee waste 
can produce the greatest number of  fruiting bodies while 
also being the most viable in terms of  weight yield. Thus, 
different mixtures of  cassava peels, coconut residue, 
and coffee waste were an effective substrate mixture 
to produce viable yields and, therefore, can be used as 
a successful substrate for oyster mushroom cultivation. 
The findings of  this study proved that cassava peels and/
or a combination of  coconut residue, and coffee wastes 
can be use as substrate option for mushroom growers due 
to its high yield potential. Aside from their nutritional, 
medicinal, and economic value, oyster mushrooms (P. 
ostreatus) may aid in agricultural waste management in 
the country. These can help bridge environmental issues 
and advance climate change response. However, further 
research on the effects of  these substrate mixtures to 
oyster mushrooms as well as its varying proportions, are 
recommended.

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