Greener
Journal of Agricultural Sciences Vol. 9(3), pp. 322-336, 2019 ISSN: 2276-7770 Copyright ©2019, the copyright of this article is retained by the
author(s) DOI Link: https://doi.org/10.15580/GJAS.2019.3.070119123 http://gjournals.org/GJAS |
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Nutrient Compositions and
Optimization of Elephant Grass (Pennisetum purpureum)
Stem to Cotton Seed Proportion for the
Cultivation of Oyster Mushroom (Pleurotus ostreatus)
at Ambo Western, Ethiopia.
Gurmessa Tesema*; Asefa Keneni
Department of Biology, College of Natural and Computational Sciences,
Ambo University, Ethiopia.
ARTICLE INFO |
ABSTRACT |
Article No.:070119123 Type: Research DOI: 10.15580/GJAS.2019.3.070119123 |
At
present more emphasis has been given to mushroom cultivation for the
nutrition and medicinal uses and Agricultural product recycling technology.
The aim of this study was to investigate the usability of Pennisetum purpureum stem as
major substrate for cultivation of oyster (Pleurotus
ostreatus) 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
collared sorghum and sterilized substrate was inoculated with 10% of the
spawn wet basis on dry basis of the substrate. The experimental design
constitutes ten treatments on the stem of elephant grass with ratios of
cotton seed wastes (T1-T10) in three replications. At T4 (70:30) the fastest
mycelial grow on the stem of elephant grass and
Slowest mycelia extension were observed on T7 (60:40). the highest fresh weight were observed on
T4 (70:30) stem of elephant were recorded 1254 g / 500g dry substrate,
highest number of fruits recorded on T5 (62) while largest cap diameter
(11.5cm) was recorded on T2 of the stem of elephant grass the highest stipe
length were recorded on T9 (3.67cm) . The highest biological efficiency
observed on T4 (250.5%) on stem of elephant grass. the highest protein
contents were recorded on T7 (36.17%) on stem of elephant grass with
supplementation of cottonseed wastes and the least one were recorded on T9 (80:20) 16.87%, on the
others treatments intermediate number of food contents were recorded. Even
though on the stem of elephant grass and the proportion supplementation of
cotton seed waste was varied for all the treatments except on T1 treatments
make 100% Pennisetum purpureum
dry stem were made. Over all, the results of the study showed that the Pennisetum purpureum stem
supplemented with cotton seed wastes can be gave highest yield and yield
related parameters of oyster mushroom. The results of the present study
implies to carry out further research on the optimization of Pleurotus ostreatus production
using two or more type of substrates tested in this experiment. |
Submitted: 01/07/2019 Accepted: 09/07/2019 Published: |
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*Corresponding Author Gurmessa Tesema E-mail: gurmesat2014@
gmail.com |
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Keywords: |
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Pleurotus species,
commonly known as oyster mushrooms, are edible fungi cultivated worldwide
especially in south East Asia, India, Europe and Africa (Mandeel et al., 2005).
China
produces 64 % of all edible mushrooms in the world and 85% of all oyster
mushrooms all over the world (Pleurotus spp.)
is also produced in China (Chang.,1999). Mushroom
production in rural communities can alleviate poverty and improve the
diversification of agricultural production (Godfrey et al., 2010).
Mushrooms
cultivation offers benefit to market gardens when it is integrated into the
existing production system by producing nutritious food at a profit, while
using materials that would otherwise be considered “waste” (Beetz
and Kustudia, 2004). This is because mushrooms
contain many essential nutrients and they are found to solve dietary related
health problems (Atikpo et al., 2008)
It is a valuable mushroom with good marketability and
is relatively easy to grow. It requires no arable land for production and the
abundant agricultural waste found countrywide offers opportunity for
production, which in turn provides a more economical and environmentally
friendly disposal system (Stamets, 2009; Olfati and Peyvast, 2012; Philippoussis
and Diamantopoulos, 2012).
There are many types of mushrooms that offer a long
list of health benefits. Among them oyster mushroom occupies 14% of the global
market and ranks third in the global trade. It tolerates a temperature of 7 -
37°C with an optimal range of 26 - 28°C and is rich in protein, fiber, iron,
vitamins and minerals (Wani et al., 2012).
According to (FAO,
2009), it is an enterprise for both men and women and it is especially an
excellent enterprise for women since it does not demand much labor and energy
for production. Mushroom production indirectly provides materials that are used
to improve the soil structure for production of other crops. Mushroom
cultivation is a useful method of environmental waste management and waste
disposal. Mushrooms are also considered a good source of protein, considering
protein content of dry mushroom, however, it is important to emphasize that the
protein content of fresh mushroom is hardly higher than 4% (Silva et
al., 2007; Bernardi et al., 2009).
Mushroom cultivation has been reported as other effective way of alleviating
poverty in developing countries (Masarirambi et al., 2011).
However,
there is no mushroom cultivation practice in the country to fill the demands of
people interested in the mushroom consumption. Those very few mushroom farms in
Ethiopia are restricted to the capital city. Some research based practices in
some parts of the country are still at the stage of trials. The current study
was, therefore, initiated to assess the suitability of differ-ent locally available substrates and their combinations for
cultivation of P. ostreatus and to estimate
yields of cultivated mushrooms on different locally available cheap substrates.
Elephant grass is very important forage in the tropics due to
its fast productivity. It is particularly suited to feed cattle and buffaloes.
Elephant grass is mainly used in cut-and-carry systems ("zero
grazing") and fed in stalls, or made into silage or hay. Elephant grass
can be grazed, provided it can be kept at the lush vegetative stage: livestock
tend to feed only the younger leaves (FAO, 2015). Elephant grass, as implied by its name, is an important source of
forage for elephants in Africa (Cook et al., 2005). However the aimed through this research to
reveal the use of inexpensive agricultural foods to grow mushrooms and
evaluates suitability of Elephants grass (Pennisetum
purpureum) for
cultivation mushroom.
Statement of the problem
Mushroom
cultivation could be a possible option to alleviate poverty and malnutrition in
developing countries (Diriba et al., 2013). The science of mushroom growing is now
confined to a few producers in the country. A number of research activities have been
carried out on the cultivation of oyster mushroom using different agricultural
waste products worldwide. Several publication have focused on the
utilization of different composition of agricultural waste products as substrate
for mushroom production (Asefa and Geda 2014b)
.However, the utilization of Elephant grass (Pennisetum
purpureum) which is grown enormously in the
swampy, waterlogged areas in our country has not been attempted so far as a
substratum for growing mushroom species.
The Elephants grass (Pennisetum purpureum)
is available at high amount in different localities such as agricultural
centers, and spring areas in Ambo University. Inside the campus of Ambo
University also there is plenty of this grass were available and left unused.
In this context this research has been initiated in order to understand the
possible utilization of this plant biomass along with cotton seed waste as
substratum for growing the oyster mushroom (P.ostreatus). It is aimed through
this research to reveal the use of inexpensive agricultural food to grow
mushrooms and evaluate suitability of Elephants grass (Pennisetum purpureum) for cultivation mushroom.
The
practice of mushroom cultivation is not well known in Ethiopia even though a
number of attempts have been made to make awareness among the community.
Several studies have been conducted in the utilization of various plant parts
as substratum for the cultivation of oyster mushroom. However, no research has
been conducted on Elephant grass as a substratum for mushroom cultivation in
Ethiopia. Therefore, the present study
was conducted to explore the possibility of using these plants for the
cultivation of the oyster mushroom. Moreover, the results of this study would
initiate more in depth research on the application of this plant for different
species of mushrooms
MATERIALS AND METHODS
The research study was carried out at Ambo
University, Ethiopia, in mushroom production center (MPC); the institution is
located about 116 km away from the capital city of Ethiopia. Geographically
Ambo University main campus, which was located at the altitude of this
institution, is 2101 meter above sea level. The Ambo city was
located at Latitude: 8°
58' 59.99" N and Longitude: 37° 50' 59.99" E.
the selected area has good climate condition to cultivate
mushroom production. Its temperature ranges from 19-29oc.
Organism and culture conditions
The fungal strain, Pleurotus
ostreatus (Oyster mushrooms) were obtained from
Micro biology Laboratory, Department of Biology, Ambo University, Ambo,
Ethiopia. The pure culture of Pleurotus ostreatus were transferred on to Potato Dextrose Agar
(PDA) prepared in the laboratory and chloramphenicol 0.2 g in 1000 ml of water.
The medium were poured into the Petri dishes and allowed to cool in under
aseptic condition in laminar flow chamber. The cooled and solidified medium
were inoculated by 1 cm×1 cm agar block of the fungal strain and incubated at
25oC. The growth of the culture and presence of contamination were
visually inspected at three days interval.
In this study, the spawn (mushroom seed) of Pleurotus ostreatus was
prepared on yellow colored sorghum grain, Wheat brain and calcium sulfate
(gypsum) in the ratio of 88:10:2, respectively (Dawit
Abebe, 1998). The required amount of sorghum grain
was weighed and soaked overnight 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 (CaSO4.2H2O) 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 ± 20C
until the substrate were fully colonized and the mycelia invasion and
contamination were inspected at five days interval. After 15 to 21 days the packet of the culture
become white due to the completion of the mycelium running and then it was
ready for inoculation on the substrates.
Substrate
preparation and Processing for inoculation.
The
substrates were prepared from Elephant grass, with the addition of cotton seed.
Cotton seed were collected from Addis Abeba city
markets and Elephant grass were collected from the Ambo University main campus.
Stem of Elephant grass was chopped (2-3 cm) and dry these substrates were
soaked in water over night to get wet and achieved 65-70 % of moisture content
(Shown in Figure 3.1 below). The next day, all these wet substrates were
separated from water and Excess water present in the substrates was drained
thoroughly and mixed with required amount of calcium carbonate (1%) and filled
in sterilizable yellow color polyethylene bags (Kurtu pestal).
The
substrates were autoclaved at 15Psi pressure at 1210C
temperatures for 1h. After cooling the sterilization substrates were
transferred to transparent polyethylene cultivation bags for easy supervision
of the growth of the mycelia and presence of contamination. Each substrate (500
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. Mycelium running rate on the substrates was observed after
7 days inoculation of spawn. It was kept in a spawn running room at room
temperature in the dark until primordial were formed. After primordial
formation, large holes were made in the polythene bag to allow normal
development of fruiting bodies.
Bags
were transferred to mushroom house under normal environmental conditions and
relative humidity (the room maintained at 85-90%) by keeping water in open
containers at different corners of the room. The cultivation bags were
irrigated using tap water every morning and evening until all flushes of Pleurotus ostreatus fruiting
bodies were harvested. Adequate ventilation was provided to prevent increased
CO2 concentration in the room by opening the door and windows of the
room for half an hour in the morning and in the evening. The fruiting bodies of
mushrooms were manually harvested at maturity which was indicated by upward
curving of the edges of the cap.
Experimental Design
Ten
treatments (T1–T10) comprising different proportions Stem of Elephant grass and Cotton seed wastes,
the combination of them are (500 g) along with 1% of lime stone (Calcium
Carbonate) on dry weight basis were used. Experiment design is a completely
randomized design with three replications per treatment, being each an
experimental unit.
Table
.1. Ratio of the Stem of elephant grass and
cotton seed in the treatments
Treatment |
(SE) Stem of Elephant grass(gram) |
(CW)
Cotton seed (gram) |
Total |
S: C Ratio
(%) |
||
T1 |
500 |
0 |
500 |
100:0 Control |
||
T2 |
450 |
50 |
500 |
90:10 |
||
T3 |
400 |
100 |
500 |
80:20 |
||
T4 |
350 |
150 |
500 |
70:30 |
||
T5 |
300 |
200 |
500 |
60:40 |
||
T6 |
250 |
250 |
500 |
50:50 |
||
T7 |
200 |
300 |
500 |
40:60 |
||
T8 |
150 |
350 |
500 |
30:70 |
||
T9 |
100 |
400 |
500 |
20:80 |
||
T10
Control |
0 |
500 |
500 |
0:100 |
||
The
performance and productivity of the mushroom were measured using the method
outlined by Mkhize et al. (2016). The following parameters were measured in order to
evaluate the performance and productivity of P. ostreatus
mushroom.
The number of days it took for the mycelia to fully colonise
the substrate as also noted from the time it first inoculated the substrate up
to a point where the mycelia full covered substrate. These
included the biological efficiency, fresh mushroom yield, duration of mycelial growth rate, duration of
primordial formation, duration of maturation of mushroom, time to fruiting
bodies, Cup diameter, and stipe length of the mushroom were measured.
The mushroom yield was calculated according
to Morais, et
al. (2000), using the equation: The main parameter used to evaluate
mushroom yield is called biological efficiency (BE). It mainly depends on the
characteristics of the material and the circumstances in which the growth
process occurs. Biological efficiency was calculated and defined as the ratio
of weight (g) of fresh mushrooms harvested to dry weight (g) of the substrate.
Biological Efficiency = Weight of fresh
fruiting bodies (g) /Weight of substrate (g) × 100.
The
moisture content is determined by measuring a material before & after the
water removed by evaporation Moisture content was determined by following the
formula
%Moisture =initial –dried/initial *100.
Where M
initial and M dried are the mass of sample before and after drying respectively
to obtain an accurate measurement of the moisture content of material
evaporation method necessary to remove all water molecules.
To about
0.7 gram of sample in a digestion flask, 1 gram of Copper Sulphate,
10 gram of Potassium sulphate and 20 ml of Sulphuric acid was added. After complete digestion the
content is transferred into a vessel. 25 ml of 0.2N Sulphuric
acid was pipette out into beaker and distillation was started. The distillate
was allowed to collect in Sulphuric acid for a known
volume and time. The collected distillate is titrated against 0.2N Sodium
Hydroxide using Methyl red as an indicator.
The
percentage of Protein was calculated. Total nitrogen
was estimated by following the standard Kjeldahl method (Chang, et al, 2003).
Where, Va = mL HCl
measured in the conical task in the distill (usually 20.00 mL)
Vb = mL NaOH used for
titration of the content in the conical flask
Na =
Normality of the HCl measured into the conical flask
Nb = Normality of the NaOH
used for titration
W = g of
mushroom powder used for the analysis
Crude protein content was obtained by
multiplying the total nitrogen value by the conventional factor 6.25. (Chang et al, 2003). The percentage of protein
in the sample was calculated by the following equation:
Crude
Protein (%) = % N X 6.25
Total
lipid was determined by slight modified method of Folch,
et al. (1957). Five gram of each
sample was suspended in 50ml of chloroform: methanol (2:1) mixture then mixed
thoroughly and let stand for 3 days. The solution was filtrated and centrifuged
at 1000 rpm by a centrifuge machine. The upper layer of methanol was removed by
Pasteur pipette and chloroform was evaporated by heating. The remaining was the
crude lipid.
One gram
of the sample was weighed accurately into a crucible. The crucible was placed
and heated first over a low flame till all the material was completely charred,
followed by heating in an oven for about 6 hours at 6000C. It was
then cooled and weighed. Then total ash was calculated as following equation (Raghuramalu et al.,
2003).
Ash content (g/100g) = weight of ash *100/ weight of sample
5 gm of mushroom sample was extracted using Petroleum ether.
The fat free material was transferred in a beaker and 200 m1 of dilute sulphuric acid was added and boiled. Whole boiling acid in
a flask is connected to reflux condenser and heated for 30 minutes. The flask
was removed and filtered and washed thoroughly with boiling water followed by
washing in boiling Sodium Hydroxide and again refluxed for 30 minutes. The
contents were filtered and washed with boiling water and finally washed the
ethanol. The residues were dried and incinerated in muffle furnaces at 660
degree Celsius and the crucible along with ash was weighed and percentage of
fiber was calculated.
% of crude
fiber = 100(Wt of crucible with before ashing- Wt of crucible after ashing)
Weight of Sample
The
content of the available carbohydrate were determined by the following equation
(Raghuramalu et
al., 2003).
Carbohydrate (g/100g sample) = [100 – (Moisture + Fat + Protein + Ash +
Crude Fiber)]
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 21versions of Statistical Package for Social Sciences
(SPSS) and the treatment mean will be compared using on (LSD).P≤ 0.05%.
The Pleurotus ostreatus was cultured on malt extract agar and
potatoes dextrose agar for 7 days at 28°C and mycelium covered the plate. It
was fully grow on plates as shown in (figure below). PDA and MEA were the
simplest and the most popular medium for growing mycelia of most cultivated
mushrooms (Chang, 1999). P. ostreatus was successfully grown on PDA and MEA. The
oyster completely covered the peteridishes after 7
days and its color and appearance looks like pure cotton. The mycelium should be white and grow out
from the tissue. If yellow, blue, green or grey mycelia form on other places on
the surface, then these are fungal contaminants (Oei
and Nieuwenhuijzen, 2005). A creamy, shiny growth
often indicates bacterial contamination (Oei and Nieuwenhuijzen, 2005). P.
ostreatus is a slow grower when it is compared to
molds and other fungi.
Spawn
production
In this
research or experiment, yellow Sorghum was inoculated by P. ostreatus
for spawn production of oyster mushroom. Sorghum based spawn took 25 days to
colonize the substrate completely (figure 3 below). The moisture content of the
sterile moist sorghum (65-70%) was found to be suitable for growth of mycelium
of oyster mushroom. The mycelium was completely colonizing sorghums and it was
completely change the color to wheat. Those fully colonized by mycelium without
any contamination of microorganisms and ready for inoculation of the
substrates, Sara, (2007).
Day’s taken mycelial
extension on stem of elephant grass
The fastest grow on the stem of elephant
grass, it takes 14th to 15th days on the T4 and
T3 treatments and the lowest mycelial
extension were recorded on T7 32 days .the other were required intermediate
days to colonize mycelial extensions from days of
inoculation stem of elephant grass. (Shown Table 2 below).
There were significant (P≤0.05) differences in the days required for
complete invasion mycelial on the substrates
receiving for different treatments. The time required for complete colonization
of the substrate by oyster mushroom was longer on stem of elephant grass
supplemented with a cotton seed waste at ratios of (40:60). (Follow table 4
below).This result was also line to, the mean value of mycelia extension
reported by Gume et
al, (2013), Mekonnen and Semira,
(2014). And Asefa and Geda, (2014 b).
Gume et
al., (2013) reported the highest mycelial running
rate was observed in substrate composed from saw dust maize comb and coffee
husk. In this study, there were slight differences on days required for
complete colonization of the substrates that received different treatments.
Also the growth was observed on the different ratios of the substrate
from stem of elephant grass and on different ratios of stem of elephant grasses
supplemented with a cotton seed wastes. The first primordial
appears after 14 days after inoculation depending upon types of substrate. The
primordial formation and number of primordial per plastic bag (substrate) was
affected by humidity, aeration and the substrate itself. Number of primordial
was highly growth or appeared on the ratio of 70:30 (T4) of the substrates on
stem of elephant grass. It indicates the growths was formed on the high ratios
of treatments stem of elephant grass with addition of cotton seed wastes and on
the other treatments at the intermediate duration of time number of primordial
were produced. But when we compare
leaf and stem of elephant grass with ratios of cotton seed wastes highly its
growth on the stem of elephant grass than on the leaf of elephant grass. (Shown in figure 4 below).
The effect
chopped stem of Pennisetum purpureum were evaluated on different treatment of the
substrates supplemented with cotton seed wastes. The number of fruit body and size of fruit
body were produced on the different treatments. The fruiting body of mushroom
was highly growth on the all treatments of substrate on stem of elephant grass
and mixed with the cotton seed wastes.
The fruiting body formed on all treatments stem of elephant grass, the
size and the numbers of fruiting body were different from substrates to
substrates of the treatments. The large fruiting body were collected on the T5
(40:60), when the ratios of treatment of the substrates were more and on the
others also T3 (80:20), on the high ratios of the substrates good quality and
size large fruiting body were collected. Follow figure below.
The number
of primordial were first observed at
the T4 (70:30) and T2 (90:10) and T3 (80:20) at the ratios of stem of the
elephant grass were more and some of the slow primordial formation were T6
(50:50) and the other treatments were observed intermediate number days of
primordial formation after mycelial colonization for
all. There were significant (P≤0.05) differences in the primordial
formation of oyster mushroom grown on stem of different Treatments. (It was
shown in the table 2 below). These longer days of initiation of primordial
formation after mycelia running may be due to slow releasing of nutrients from
the both leaf and stem of the substrates as compared to other treatments, for
example, wheat straw and rice straw on which much of research work has been
done on this mushroom species. The observed result was near line to similar
with Ashraf (2013). Ashraf et al.,
(2013) reported that all the treatments they tested showed 3.73 to 5.13 days
for primordial initiation after mycelia running.
There were
not significant (P≤0.05) differences in the maturation formation of
oyster mushroom grown on stem different Treatments. On the second substrates
stem of elephant grass more number of days taken on the T6 (50:50) 49th
days taken to harvest and T7 (60:40) 47.5th days were recorded and
the first one harvested were on the T4 or the shortest number days observed and
others were required intermediate number of days to harvested the fruiting body
of mushroom. But when we compare both of the treatments the second treatments
stem of elephant grass with addition of cottonseed wastes take some number of
days difference to harvested mushroom than the first treatments on leaf of
elephant grass with addition of cotton seed wastes to harvested mushroom.
(Shown on table 2 below).The period of primordial to maturation of mushrooms in
this study, the shortest mean duration was 5 days and the longest was 10 day
throughout the treatment substrates to the treatment of substrates. (Follow the
table 4.1 and 4.2 bellow). This near agrees with the range of maturation
period (3.29 to 4.33) of Pleurotus species reported by Islam et al., (2009).
Table. 2. Days
for the emergence of the various growth parameters of P. ostreatus on Elephant grass stem
substratum
Treatment |
Mycelial extension |
Primordial formation |
Maturation of mushroom frist
harvested |
T1 |
18 |
21 |
28 |
T2 |
16 |
18 |
25.5 |
T3 |
15 |
18 |
24 |
T4 |
14 |
16 |
20.66 |
T5 |
18 |
22 |
31 |
T6 |
30 |
46 |
49 |
T7 |
32 |
37 |
47.5 |
T8 |
30 |
34 |
39 |
T9 |
19 |
24 |
33.33 |
T10 |
10 |
16 |
25.7 |
MEAN |
19.89 |
24.9 |
28.67 |
STDEV |
7.5 |
9.89 |
13.5 |
There were
not significant (P≤0.05) differences on the Fruiting body of oyster
mushroom grown on stem elephant grass with addition of cottonseed wastes. On
the stem of elephant grass with the addition of cotton seed wastes the average
number of fruiting body formed was averagely 39.4.The highest number of fruits were observed on
the stem of elephant grass on the treatment T5 (60:40) counted 62, and the
least number of fruits was observed on stem of elephant grass on the treatment
one of substrate T1 (100:00), 27 were counted and High number of aborted were
recorded on the T10 and least number of aborted were observed on T2 and T6. (Shown on figure 7 below). The results of the study were
found less than the number of fruiting bodies with the previous findings of Bhuyan, (2008), and Sarker,
(2004). According to those authors the highest average number of fruiting
body/packet was observed in the treatment T3 (122.3) and the lowest average
number of fruiting body /packet was in the treatment T2 (76.0).
There were not significant (P≤0.05)
differences in the cup diameter and Stipe length of oyster mushroom grown on
stem of different Treatments. The number of cup diameter observed from the
different ratios of the substrates was different from treatment to treatments
of substrates and the averagely mean were 8.2393 respectively on Stem of
elephant grass supplanted with a cotton seed wastes. On this treatments on the stem of elephant grass with the ratios
of cotton seed wastes the largest cup diameter were observed on the T2
(11.5cm), T10 (11.8cm) on control of the experiment, and the least cup diameter
was measured on the T1 (7cm). (Shown on figure 6 below) .the others were measured
intermediate to each other. This is much greater than the pilus
diameter reported by (Gume et al., 2013). According to these authors, mean pileus
diameter of mushrooms ranged from 3.8 to 5.2cm.
The
highest Stipe length were recorded on the T4 (4cm) and the least one were
recorded on the treatment T2 (2.5cm) on stem of elephant grass with a ratios of
cotton seed wastes. (Shown on figure 6 below). The
result of this study were similar line with the study of Oseni
et al. (2012) observed Stipe length
of oyster mushrooms ranging from 39.4–59.5 mm (3.94–5.95cm) on fermented
sawdust substrate supplemented with different wheat bran levels. The result
observed in this study similar to the result reported in literature by from (Mdconline, 2013). On average, the cup diameter ranges
between 2-15 cm and stalk length is around 4 cm (Mdconline,
2013). When the number cup diameter was measured largest in number the length
of the Stipe were less measured in number. The Stipe length of the samples
collected from different treatments show significant variation.
Figure. 6. Cup
diameter and Stipe length of Oyster mushroom on different proportion of stem of
elephant grass with cotton seed wastes
Figure. 7. Number
of bunches, Fruits and Aborted of Oyster mushroom on different proportion of
stem of elephant grass with cotton seed waste
Yield
of mushroom per flush on stem of the substrates
The gram
weight of flush on the stem of elephant grass with a ratios of cotton seed
waste the highest one were observed on the T4(1254) and T1(580) was show least
in gram when we compare with in a treatments. Also high yield were observed on
the control T10 (1228). (Shown on table 3 below). The comparation between substrates stem of Pennisetum
purpureum the
yield observed in gram were almost similar to each other. In all
treatments from the cycle one to 2nd to 3rd and to 4th
the yield were reduced and the cycle were completed within 70 days. Kimenju et al.
(2009) reported that yields of mushroom in different substrates slightly
declined from the first flush to the successive harvests. Our observation on
the different harvest is in line with reports in the literature. Ashraf et al., (2013) reported that the
different treatments vary in the amount of mushroom yield harvest at different
flushes and at each successive harvest, the amount of the yield declined.
Table. 3. Yield
Fresh mushroom on different treatment stem of elephant grass
Treatments |
1st Flush |
2nd Flush |
3rd Flush |
4th Flush |
Total |
T1 |
205 |
196 |
99 |
80 |
580 |
T2 |
370 |
300 |
190 |
108 |
968 |
T3 |
450 |
330 |
270 |
180 |
1230 |
T4 |
500 |
380 |
204 |
170 |
1254 |
T5 |
420 |
260 |
210 |
121 |
1011 |
T6 |
370 |
245 |
199 |
175 |
989 |
T7 |
350 |
224 |
165 |
99 |
838 |
T8 |
340 |
255 |
203 |
185 |
983 |
T9 |
336 |
197 |
115 |
78 |
726 |
T10 |
530 |
308 |
210 |
180 |
1228 |
The effect of different treatments on
biological efficiency of oyster mushroom showed significant (P≤0.05)
differences on stem of elephant grass with addition of cotton seed wastes. The BE were recorded on this substrate
treatments of stem of elephant grass supplemented with a cotton seed wastes the
highest biological efficiency were recorded on T4 (250.8%) at the ratios of
70:30 and least one were recorded on T1 (116%) at the ratios of 100:00 stem of
elephant grass. Intermediate numbers of biological efficiency were recorded on
the others and the highest one were also recorded on control T10 (245.6%) at
the ratios of 100% cotton seed wastes. (Figure below 8).biological efficiency
obtained in this study were compared to the sawdust reported by Shah et al., (2004)reported
that B.E. remained between
21.05-64.69% when it was
cultivated on different
substrates it was not related the finding of this study. Nunez
and Mendoza (2002) it’s reported the biological efficiency values were varying
from 50.8 to 106.2 % in Pleurotus ostreatus on
different substrates were less than the present studies. The observed results
were not related to the finding of Patra and Pani, (1995), who reported the biological efficiency
(50-75%) of Pleurotus species grown on most of agro
industrial residues, namely; corncobs, various grasses and reed stems, vine
shoots, cottonseed hulls and sugarcane baggase. In
this finding, biological efficiency was indicated for comparison between
treatments of the substrate in which, the most effective substrate in
bioconversion to fresh fruiting bodies for cultivation of Pleurotus species was no more different number observed. The highest number
of biological efficiency recorded on leaf of elephant grass followed by stem of
elephant grass with addition of different ratios of cotton seed wastes. Both substrates
have more cellulose were to supported the fast Mycelial
growth during cultivation of Pleurotus species.
Nutritional analysis of P. ostreatus mushroom
Also the moisture contents of the
second treatments on the stem of elephant grass with the addition of cotton
seed wastes no more difference from the first treatments on the leaf of
elephant grasses. The moisture contents of stem of elephant grass were between
87%-92%. The highest number of moisture was observed on the treatment T6 and
least in number of moisture was observed on the T3, at the ratios of 80:20. The
result was almost near similar line to Alam; et al., (2007) reported 87–87.5%
moisture for existing Pleurotus spp. in Bangladesh and according to Moni, et al,
(2004). Its similar result was also
observed .The moisture percentage of Oyster
Mushrooms grown on different substrates observed in this study are
supported by earlier studies by Moni et al., (88.15-91.64%) and Alam et al (87-
87.5%). Generally, fresh Pleurotus
mushroom contain 85-95% moisture (Khan, 2010). The moisture content
all different composition of the substrates were followed in the table below
(Table.4 below.).The interaction between the types of growth substrate or
substrate combinations and their types had significant (p≤0.05)
difference on moisture contents of mushrooms on stem of elephant grass
supplemented with a cotton seed wastes
Pleurotus ostreatus grown on
the stem of elephant grass on different treatments were significant
(p≤0.05) different from one another. The content of protein observed on
the stem of elephant grass with addition of different ratios of cotton seed
wastes were tested, the highest protein contents were observed on the treatment
T7, (36.17%) when the ratios of 40:60 that means at the highest ratios of
cotton seed wastes and the least one were observed at the treatment
T9,(16.87%). the contents of the others treatment were inter mediate numbers on
the stem of elephant grass and it’s also recorded on control of the treatments
T10 ,(23.35%) (Shown in Table 4. below).The result of the present study similar
line with the studies of Chang et al.,
(1981) who reported that the fruit bodies of Oyster Mushrooms contained
26.6-34.1% protein. The crude
protein results of this study is close to the finding of Hassan and Medany, (2014), who reported 26.83% of crude protein
for P. ostreatus. The results are all most
similar to Breene, (1990) who reported values of
crude protein content ranging from 19-39%. Protein content of
mushrooms depends on
the composition of
the substratum, size
of pileus, harvest
time and species of mushrooms (Bano and Rajarathnam, 1982).
Protein content of the mushrooms
has also been reported
to vary from flush
to flush (Crisan and
Sands, 1978). Haddad and Hayes, (1978) indicated that
protein in A. bisporus
mycelium ranged from 32
to 42% on
the dry weight basis. Ogundele et
al., (2017) observed that the protein
content varied when culture on different substrate which related to the
nutrient composition of the substrates used.
The content of carbohydrate observed
on the stem of elephant grass with addition of different ratios of cotton seed
wastes were tested, the highest protein contents were observed on the treatment
T7, (57.78%) when the ratios of 40:60 that means at the highest ratios of
cotton seed wastes and the one least
contents carbohydrates were observed at the treatment T9,( 42.9%). the
carbohydrate contents of the others were followed on the stem of elephant grass
.the inter mediate numbers were observed on the others and T10 on the control
of the experiment,(45.18%) of carbohydrate were observed. (Shown
in Table 4. below). There were significant (P≤0.05) differences on
the carbohydrate contents of oyster mushroom grown on stem of different
Treatments. The carbohydrate content is in
agreement with the report that Carbohydrates constitute the prevailing
component of mushroom dry matter; usually about 50-60% (Deepalakshmi
and Mirunalini, 2014).The carbohydrate contents of
both of them were no more different from each other. The results observed were
near line similar to the result reported by (Deepalakshmi
and Mirunalini, 2014).Also the results of this study
were match to the study of Alam et al., (2007) who found 39.82-42.83% of carbohydrates in Pleurotus spp.
The content of fat observed on the
stem of elephant grass with addition of different ratios of cotton seed wastes
were also tested, the highest fat contents were observed on the treatment T9,
(4.31%) when the ratios of 80:20 that means at the highest ratios of cotton
seed wastes and the 2nd were observed at the treatment T3 (4.26%) on the ratios
of 80:20 and least one were recorded on T7, (2.75%) and T10 (3.84%) on control
experiment. (Shown in Table 4. below).
The fat contents of both of them were no more different from each other. The result
of this study was similar result With Alam e t al., (2007), its reported Pleurotus
mushroom ranging between (4.30-4.41).
The result observed were similar line to (Khan, 2010). Its reported Pleurotus
mushroom contain 0.5-5% of fats. The results
obtained in this study were close to that obtained by (Reguła
and Siwulski, 2007), who reported 2.66% crude fat for
dried oyster mushroom. There were not significant (P≤0.05)
differences on the fat contents of oyster mushroom grown on stem different
Treatments.
Crude
fiber (CF) on stem of elephant grass
The content of Crude fiber observed on
the stem of elephant grass with addition of different ratios of cotton seed
wastes were tested, the highest Crude fiber contents were observed on the
treatment T6, (20.67%) when the ratios of 50:50% that means on the equal ratios
of the substrates. and the least one were observed on the T8, (17.5%).On the
control experiment T10, (18.31%) were recorded. (Shown in Table 4. below).There were
significant (P≤0.05) differences on the fiber contents of oyster mushroom
grown on stem different Treatments.
The crude fiber contents of both of them were no more different from each
other. According to Teklit
(2015) has compared the nutritional composition of cultivated mushrooms in
Ethiopia and found that the crude fiber content varies from 18.23-29.02% .it
also similar result observed in these studies. The results observed were near the result gained by (Kalac. et al.,)
having reported that about 4-9% and 22-30% for soluble and insoluble fiber,
respectively (Kalac. et al., 2009).
Crude
Ash (CA) content on stem of elephant grass
The content of ash observed on the stem of
elephant grass with addition of different ratios of cotton seed wastes were
tested, the highest ash contents were observed on the treatment T5, (15.92%)
when the ratios of 60:40 that means at the highest ratios of stem of elephant
grass and the least one were observed on T6, (11.73%) and T10 (8.41%) on the
control experiments. (Shown in Table 4 below).There were
significant (P≤0.05) differences on the Ash contents of oyster mushroom
grown on stem different Treatments. The
findings of the study were supported by the study of Khlood-Ananbeh,
et al., (2005). Who reported ash
contents were moderate in the fruiting bodies. Alam et al., (2007).Reported 8.28 - 9.02% of
ash in Pleurotus spp its less than the present study. In Pleurotus Florida, Teklit,
(2015) observed 9.41% ash content which less than the present study on those
grown on elephant grass substratum.
The ash contents of both of them were different from each other the present
observed on stem of elephant grass with addition of cotton seed waste was
higher than the leaf of elephant grasses.
Table. 4.The effect of additives or proportion of stem
substrates on mushroom production
T/t.2. |
Moisture (db) |
Crude
Protein (db) |
Crude fat (db) |
Crude
Fiber (db) |
Crude Ash(db) |
Carbohydrates (db) |
T1 |
89.25 |
26.78 |
4.19 |
19.533 |
14 |
53.753 |
T2 |
88.41 |
25.1 |
3.35 |
18.88 |
12.25 |
47.99 |
T3 |
87 |
21.65 |
4.26 |
17.53 |
13.27 |
43.71 |
T4 |
89.833 |
18.66 |
3.34 |
19.83 |
12.41 |
44.07 |
T5 |
89.66 |
24.08 |
3.84 |
17.65 |
15.92 |
51.15 |
T6 |
90.27 |
21.01 |
3.31 |
20.65 |
11.73 |
46.97 |
T7 |
88 |
36.17 |
2.73 |
18.68 |
12.2 |
57.78 |
T8 |
90.5 |
25.05 |
2.79 |
17.5 |
12.32 |
48.16 |
T9 |
90.5 |
16.87 |
4.31 |
18.31 |
12.91 |
42.9 |
T10 |
92 |
23.28 |
3.19 |
18.31 |
8.41 |
45.18 |
There were significant (P≤0.05) differences on the
nutrient contents of oyster mushroom grown on stem of different Treatments
Cultivation of edible mushroom has been
considered as an additional practice of food production and contributes in the
struggle for food security and solving the problem of malnutrition in
developing countries. The use of Pennisetum purpureum stem as
a major substrate with the supplementation of cotton seed waste has not yet
been tested in the production of oyster mushroom in Ethiopia. The present study
reveals the usability of Pennisetum purpureum stem and cotton seed wastes with respect to gave the highest yield, yield para
meter, biological efficiency and nutrient of the oyster mushroom. Based on the
results of the study the following conclusion was made: The highest total yield
was collected on T4 (1254g/500g) of dry substrates on the stem of elephant
grass and Highest biological efficiency was recorded on T4 (250.5) stem of
elephant grass and on all the other treatments intermediate results were
recorded. The highest stipe length (4.5cm) on T6. And
highest number of fruits (62) was recorded on T5 While the largest cup diameter
(11.5cm) on T2 and the highest stipe length (3.67cm) on T9 of the stem of
elephant grasses the major substratum. The highest protein contents were
recorded on T7 (60:40) 36.17% and the protein contents were recorded on T9
(80:20) 16.87%.
I would like to thank most sincerely my major
Advisor Dr. Asefa Keneni,
for his continuous guidance, material provision, constructive comments and
suggestions from the beginning to the final preparation of this research. His
dedication and professionalism is always appreciated. Also I would like to
thank the Biology and Chemistry
departments of Ambo University, for the supply of necessary chemicals and equipments to conduct the laboratory analysis.
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Cite this Article: Gurmessa, T; Asefa, K (2019). Nutrient Compositions and Optimization
of Elephant Grass (Pennisetum purpureum)
Stem to Cotton Seed Proportion for the Cultivation of Oyster Mushroom (Pleurotus ostreatus)
at Ambo Western, Ethiopia. Greener
Journal of Agricultural Sciences 9(3): 322-336, https://doi.org/10.15580/GJAS.2019.3.070119123 |