Oyster Mushroom (Pleurotus ostreatus) Cultivation Using Sawdust and Different Organic Manures - Asian Food Science ...
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Asian Food Science Journal
20(5): 67-74, 2021; Article no.AFSJ.67355
ISSN: 2581-7752
Oyster Mushroom (Pleurotus ostreatus) Cultivation
Using Sawdust and Different Organic Manures
O. A. Orngu1*, I. E. Mbaeyi-Nwaoha1, B. O. Unagwu2 and V. E. Etim3
1
Department of Food Science and Technology, University of Nigeria, Nsukka, Nigeria.
2
Department of Soil Science, University of Nigeria, Nsukka, Nigeria.
3
National Biotechnology Development Agency, South East Center, University of Nigeria,
Nsukka, Nigeria.
Authors’ contributions
This work was carried out in collaboration among all authors. Author OAO designed the study,
performed the statistical analysis, wrote the protocol and wrote the first draft of the manuscript.
Authors IEMN and BOU supervised the study. Author VEE managed the literature searches.
All authors read and approved the final manuscript.
Article Information
DOI: 10.9734/AFSJ/2021/v20i530299
Editor(s):
(1) Dr. Chouitah Ourida, University of Mascara, Algeria.
Reviewers:
(1) Hari Om, India.
(2) B. N. Hazarika, College of Horticulture and Forestry, India.
Complete Peer review History: http://www.sdiarticle4.com/review-history/67355
Received 07 February 2021
Accepted 15 April 2021
Original Research Article
Published 23 April 2021
ABSTRACT
Aims: The aim was to evaluate the performance of cultivating oyster mushroom (Pleurotus
ostreatus) using sawdust and different organic manures.
Study Design: The experimental design used was the complete randomized design (CRD) and
the Data obtained was subjected to Analysis of Variance (ANOVA) followed by Turkey’s Least
Significant Difference(LSD) test to compare treatment means; differences was considered
significant at 95% (P≤0.05) (SPSS Version 21 software).
Place and Duration of Study: National Biotechnology Development Agency, South East Center,
University of Nigeria, Nsukka between September 2019 and November 2019.
Methodology: Saw dust was prepared as substrate with different organic manures and coded SD,
Saw dust; SDP, Sawdust+ Poultry; SDC, Saw dust + cow dung; SDD, Saw dust+ pig dung) in the
ratio of 100:0 and 50:50 respectively to cultivate oyster mushroom (Pleurotus ostreatus).
Subsequently, the rate of growth, time of harvest, yield and average weight were recorded and
proximate composition determined using standard methods.
_____________________________________________________________________________________________________
*Corresponding author: Email: africaorngu@gmail.com, africaorngu@yahoo.com;
Orngu et al.; AFSJ, 20(5): 67-74, 2021; Article no.AFSJ.67355 Results: Oyster mushroom harvested showed Moisture content ranged (4.63-7.14%), ash content (4.84-6.77%), crude fat (0.98-3.28%), fiber (16.02-18.23%), protein (19.27-33.41%) and carbohydrate (38.18%-48.89%). Average weight yield was highest in saw dust substrate (10.2g) with total yield (980g) and least average weight (7.5g) and total yield (105g) in sawdust and poultry litters substrate. Oyster mushroom from all the substrate differed significantly (p
Orngu et al.; AFSJ, 20(5): 67-74, 2021; Article no.AFSJ.67355
Table 1. Substrate formulation for oyster the method described by Okeke et al. [9]. The
mushroom cultivation fresh mushroom was cleaned properly to remove
dirt and damaged portion. The fresh mushroom
Substrate Ratio was blanched in hot water at 32°C for three
SD 100:0 minutes, which contained 3% salt and 0.01%
SDP 50:50 citric acid. Then, water was drained, and the
SDC 50:50 mushroom dried in an oven at temperature of
SDD 50:50 105°C for 3 hours. The dried mushroom was
Key: SD, Saw dust; SDP, Sawdust+ Poultry liters; milled in the laboratory using hammer mill and
SDC, Saw dust + cow dung; SDD, Saw dust+ pig dung passed through a 60-inch mesh sieve (British
2.2.1 Sterilization Standard Screen) and packaged in a low-density
polyethylene bag, stored in the refrigerator (4 °C)
The bagged substrates in 10×12 polyethylene until required for use.
were sterilized in autoclave for 15 minutes prior
to inoculation at the temperature of 121°C to
eliminate any microbial contamination and
allowed to cool overnight [8].
2.2.2 Inoculation of substrate
Inoculation was done after sterilization
aseptically into the sterilized bags of the
substrate. Ten grams (10g) of spawn was
weighed and inoculated to each substrate and
tied up and taken into a dark room for mycelium
colonization/ growth at 25°C - 30°C and 90%
relative humidity. Within seven days, mycelium
colonization or spawn run was observed. The
spawn run time depend on the size of the bag,
amount of spawn used, the strain used and the
temperature.
2.2.3 Fructification
After 25 days, substrate bags that were ramified
completely were moved to mushroom growth
chamber for photosynthesis to enable sprouting
and fructification. Tiny holes were created on the
polyethylene bags using a wire. The polyethylene
bags were opened and watered with 3 ml of
water each using different syringes daily to avoid
cross-contamination. Environmental conditions
such as temperature, ventilation, light and
humidity are necessary for fructification.
2.2.4 Harvesting/Packaging Fig. 1. Flow chart for the preparation of
mushroom powder
Fresh mushroom was harvested on maturation Source: Okeke et al. [9]
was carefully by hand from the substrate bags
after 31 days of inoculation. The harvested 2.3 Analysis
mushroom fruits were weighed, recorded and
oven dried at 45oC, to ensure phyto-chemicals 2.3.1 Determination of moisture content
are not lost.
2.2.5 Processing of mushroom powder This was determined in triplicate by hot air oven
method in an indirect distillation method.
The oyster mushroom (Pleurotus ostreatus) According to AOAC [10] method, three grams (3
powder was processed in the laboratory using g) each of the product sample was weighed (W 2)
69
Orngu et al.; AFSJ, 20(5): 67-74, 2021; Article no.AFSJ.67355
using a digital balance into a cleaned, dried, transferred into a pre-weighed evaporated dish.
cooked and weighed crucible (W 1). The sample, The petroleum ether was recovered by
which was in the crucibles, was transferred into a evaporation using a bath and the remaining fat in
hot air oven at 150ºC and dried for 2 hours. The the flask was dried in the oven at 80ºC for 30
crucible was weighed periodically until constant minutes and cooled in a desiccator and finally
weights (W3) was obtained. The percentage of weighed using digital balance. The difference in
moisture was calculated as follows: the weight of the empty flask and the flask with
oil (C) gave the oil content was calculated as
% Moisture = W 2- W3 x 100 percentage fat content:
W 2-W1 1
Where; % Crude fat = C − B x 100
A 1
W1 = Initial weight of empty crucible;
W2 = Weight of crucible + food before drying A = Weight of sample
W3 = Final weight of crucible + food after drying; B = Weight of empty flask
% Total solid (dry matter) = 100 - % moisture C = Weight of flask + oil
2.3.2 Determination of crude fibre 2.3.4 Determination of crude protein
The crude fibre was determined using the The crude protein content of the sample was
method described by AOAC [10]. Two grams (2 determined by the semi-micro Kjeldahl technique
g) of each of the sample (W1) was weighed on a as described by AOAC [10]. One gram (1.0 g) of
digital balance, boiled underreflux for 30 minutes the sample was put into a Kjeldahl flask and 3.0
with 200 ml of a solution containing1.25 g of g of hydrated cupric sulphate (catalyst) was
H2SO4 per 100 ml of solution which was put into added in the flask. Then, 20 ml of anhydrous
a 250ml beaker. Then, filtered through a muslin sodium sulphate and 1.0 g of concentrated
cloth on a fluted tunnel and heated with hot water tetraoxosulphate VI acid (H2SO4) was added to
for 2 times. The residue was transferred into a digest the samples, which was topped and
250 ml beaker and boiled for 30 minutes with 200 swirled. The flask and liquid was clear and free of
ml of a solution containing 1.25 g of carbonate coloration. The clear solution was cooled and
free of NaOH per 100ml. The final residue was made up to 100 ml with distilled water and a
then filtered through a muslin cloth and dried digest of 5 ml was collected for distillation. Then,
using a hot-air oven (GallenKamp oven, United 5 ml of 60% sodium hydroxide solution (NaOH)
Kingdom). The final residue was ashed in a was put into the distillation flask and distilled for
furnace and put into the desiccator to cool and some minutes. Boric acid indicator absorbed the
weighed (W3) using a digital balance; ammonia which was distilled off and this was
titrated with 0.1 ml hydrochloric acid (HCl). The
% Crude fiber = W2-W3 x 100 titer value or the end point at which the colour
W1 changes from green to pink was taken. The
crude protein was calculated as:
W1 = Weight of the sample
W2= Weight of the sample before ashing Percentage crude protein = 0.001410 x 6.25
W3 = Weight after ashing x 25 x T x 100
Where: T = Titre value
2.3.3 Determination of crude fat
2.3.5 Determination of ash content
Crude fat was determined using Soxhlet
extraction method as described by AOAC [10]. A Ash was determined in a muffle furnace as
250 ml clean boiling flask (B) was dried in an described by AOAC [10]. Crucibles were heated
oven at 100ºC using hot air oven. Three gram (3 in an oven for five minutes to eliminate moisture,
g) of the sample (A) was weighed (using a digital cooled in desiccators and weighed. Two grams
balance, Model number: No.T 320N) into thimble. of the sample was amassed into a crucible,
The thimble and its content were placed in the heated over a Bunsen burner to char after which
extraction apparatus and extraction was with complete ashing is conducted in a muffle furnace
o
ethyl ether in Soxhlet extractor for 6–8 h at a at about 600 C for 6 hours. The heating
condensation rate of at 3–6 drops per second. continued until whitish grey ash was left in the
During this process, the fat was extracted and crucible. The crucible was removed from the
70Orngu et al.; AFSJ, 20(5): 67-74, 2021; Article no.AFSJ.67355
furnace, cooled in a desiccator and re-weighed to 3.2 Oyster Mushroom Fructification,
obtain the weight of ash. Maturation and Harvest Time
Calculation: Pre-mordial formation (fruiting bodies) was first
observed after 3 days in the open mushroom
Percentage ash = chamber in substrate SD and SDD. This was 28
weight of crucible +ash − weight of crucible x 100 days after inoculation with substrate SDC and
Weight of sample substrate SDP just sprouting with no fruiting
bodies. This could be attributed to the nutrient
2.3.6 Determination of carbohydrate content composition of the organic manures used with
saw dust.
The determination of the carbohydrate content of
the product was obtained by difference as The first flush which is matured fruiting bodies
described by Ihekoronye and Ngoddy [11]. This was harvested firstly in SD and SDD. This was
was done by subtraction of the sum of moisture, after 31 days of inoculation and six days of
ash, protein, fibre and fat from the total weight of removal from the dark room to the open
100. mushroom chamber. Fruiting was just recorded
in substrate SDC and substrate SDP. It took 38
% Carbohydrate = 100 – (% moisture + ash + days after inoculation and 13 days in the open
protein + fibre + fat) mushroom chamber to harvest the first flush in
substrate SDP. Meanwhile substrate SDC took
2.4 Experimental Design and Statistical 45days after inoculation and 20 days in the open
mushroom chamber to harvest the first flush. The
Analysis
time from inoculation to harvest indicate the
performance of saw dust along the different
The Experimental design was based on
organic manures used as substrate for growth of
complete randomized design(CRD) and data
oyster mushroom. The growth performance in
analyzed using one-way analysis of
substrate SD and SDD can be attributed to the
variance (ANOVA) and mean separation by
chemical composition. This study agreed with
Duncan’s New Multiple Range Test. Statistical
Tsegaye and Tefera [13] who observed
Package for Service Solution (SPSS) version 21
that the number of fruit bodies recorded is
software was used and significance accepted at
related to their mycelia colonization. The
pOrngu et al.; AFSJ, 20(5): 67-74, 2021; Article no.AFSJ.67355
poultry litter) had only one(1) flush each. This is ash content of 4.91% with 6.45% in substrate
as shown in Table 2. SDD. The oyster mushroom samples cultivated
on the substrate differed significantly (pOrngu et al.; AFSJ, 20(5): 67-74, 2021; Article no.AFSJ.67355
Table 3. A proximate composition of oyster mushroom grown on different substrate
Substrate Moisture Ash Crude fat Crude Fibre Protein Carbohydrate
% % % % % %
a d a a d a
SD 4.63±0.04 6.77±0.02 0.98±0.28 16.02±0.01 33.41±0.01 38.18±0.02
SDP 7.14±0.01d 4.84±0.01a 3.28±0.01d 18.14±0.01c
20.45±0.07b 46.15±0.07c
c b c d a d
SDC 6.30±0.01 4.91±0.00 2.39±0.01 18.23±0.01 19.27±0.01 48.89±0.00
SDD 5.22±0.00b 6.45±0.00c 1.86±0.00b 17.10±0.01c
26.92±0.07c
42.43±0.02b
Key: SD, Saw dust; SDP, Sawdust + Poultry liters; SDC, Saw dust + cow dung; SDD, Saw dust + pig dung
significantly (pOrngu et al.; AFSJ, 20(5): 67-74, 2021; Article no.AFSJ.67355
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© 2021 Orngu et al.; This is an Open Access article distributed under the terms of the Creative Commons Attribution License
(http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium,
provided the original work is properly cited.
Peer-review history:
The peer review history for this paper can be accessed here:
http://www.sdiarticle4.com/review-history/67355
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