The suitability of different combinations of locally available agro-west substrates for cultivation of Pleurotus ostreatus PPT .pdf

Mushroom has so many health benefits and use for enviromental friendly

1. JIMMA UNIVERSITY
COLLEGE OF NATURAL SCIENCES
POST GRADUATE STUDIES
DEPARTMENT OF BIOLOGY
























Comparison of Suitability of locally Available Substrates for cultivation
of oyster mushroom (pleurotus ostreatus) in Jimma, Oromia, Western
Ethiopia.in 2016






By:Takele Shiferaw Gerba
By: Takele Shiferaw Gerba

2. Introduction
Back ground
 Food insecurity, or not having the resources to obtain enough, safe,
nutritionally adequate food to support an active, healthy life, is a
significant public health issue.
 Poor people may be food secure if they compensate for their limited
financial resources with high levels of food literacy, knowledge and
experience about how to obtain and prepare cheap, high quality food
(Borch and Kjaernes, 2016).


2

3. Back ground

Doe to rapid industrialization, the amount of waste materials has increased and
utilization of these wastes is very important for government economy and
natural balance (Yildiz et al., 2002)
Producing cultured mushrooms can be one suitable solution to this problem.
Mushrooms contain many essential nutrients and they are found to solve
dietary related health problems.
•Mushrooms considered as delicious and nutraceutical food for the human health
but, still could not properly advertised in Ethiopia (Yenealem, et al.,2013).
• That very few mushroom farms in Ethiopia are restricted to the capital city.





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4. Back ground...
Generally mushrooms have a broad spectrum to serve as:
Decomposer ,waste management agent and Bioremediation no adverse impact.
Helps to reduce poverty, create job opportunity and source of income (Masarirambi et
al., 2011).
Have medicinal value (Boa, 2004).
Nutraceuticals responsible with their antioxidant, antitumor, antimicrobial
properties.
Mushrooms are consumed to prevent cancer and cardiac diseases, to improve
blood circulation and to reduce cholesterol ( free from cholesterol).
Used for physical and emotional stress, gastric ulcers & diabetes.

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5. Back ground...
High protein and low fat energy contents
The crude protein levels in P. ostreatus were reported (24.83–
27.23%) (Ashraf et al., 2013)
In general eaten as meat substitutes and flavoring agents.
oHigh productivity per unit area &Short life cycle.
oRecycling of elements & limit global warming
o As a sound environmental protection strategy
(Eswaran & Ramabadran, 2010).

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6. Objectives of the Study
General objective
To Compare locally available agro-west substrates on oyster
mushroom cultivation in Jimma University, southwestern Ethiopia
Specific objectives
To assess the suitability of different combinations of locally
available agro-west substrates for cultivation of Pleurotus ostreatus
To compare yield parameters of oyster mushroom on agro-west
substrates.
 To identify the role of environmental factors on cultivation of
Pleurotus ostreatus species.
To initiate sustainable process of conducting experiments and
mushroom cultivation (p.ostreatus species).

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7. 3. Materials and methods

. Description of Study Site and period:
The study was conducted at Jimma town, located 353 km
southwest of Addis Ababa, the capital city of Ethiopia
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8. 3.1. Culture source& preparation
Pleurotus ostreatus smushrooms were obtained from A.A.
University.
Pure cultures were transferred into sterile (PDA) plates and slants
aseptically under lamina airflow
Stored around temperature ( 25
o
C) with careful sterilization of
incubator using alcohol per day
 Whereas non-contaminated pure plate cultures were directly
used for the spawn preparation
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9. 3.2. Spawn production

Sorghum were soaked in tap water overnight for 40 h.
•Excess water was drained off &
• supplemented with (1 %) of calcium carbonate (CaCO
3).
•Sorghum grains were inoculated by transferring 2-pieces in a single bottle
approximately 1cm
2.

•The bottles were kept temperature 25, 30, 35 and 40°C.







A
B
C
D
E
9

10. 3.3. Mushroom growing room


•3.3. Mushroom growing room
Jimma University Post Graduate Micro Biology Laboratory a 3
x 3m room was prepared aseptically :
Washing the Walls, Shelves and Floor before 48h
dark environment for incubation of mushroom bags.
 Five plastic containers holding 20L


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11. 3.4 .Main substrates
3.6.1. Source and collection of substrate
The substrates used in this study can be considered practical and
economically feasible due to their availability throughout the year at
no cost in large quantities in western part of Ethiopia.
Five different types of main substrates namely:
 Saw dust of (Cupressus lusitanica ) locally called Tid
Cow dung,
Teff straw (Eragros tisteff ),
Corn cobs (Zea mays),
 Chat (Khat) left over(Catha edulis) were used as main materials.

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12. 3.5.1. Combinations and Substrate preparation

The main substrates :
Were randomly Designed and triplicate in different proportion
(1:0.75,1:0.50,1:0.25 ratio each) with control treatment bags
The substrates were chopped to < 1 cm pieces
 Soaked in water over night (12h) & excess water was drained
off.
The squeeze method was used to determine the moisture content
(Bonginkhosi et al., 2012)).


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13. 3.5.2. Combinations and Substrate
preparation…
The following additives were added, alternatively
 3% gypsum (CaSO
4) chalk (CaCO
3 ) was added to 5% Maize bran.
The buffer and maze bran were mixed separately (Tesfaw et al., 2015).

105 bags
2-3h
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14. 3.6.1. Incubation and control of the
environment condition
All treatment bags were properly managed to observe effect of :
 Temperature (25 ºC), RH (80 to 90%), contaminations ,pore
size and pests that could happen.
Sufficient light & ventilation (O
2 concentrations)
 left open windows at night and 3 times per day
3.6.2. Mycelial invasion measurement: measured in weekly
basis following the method used by Rajapaks et al. (2007) and the
results were recorded


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15. 3.8.Contamination control
Sufficient sterilization with aseptic techniques was applied to
remove organisms in the substrate:
 During inoculation and cultivation the invention of pathogenic
fungi and bacteria via tools, shoes, clothes and hands were
saved by stem sterilizing, chemicals and
 The grower used reserve (safety) dressing kept in growing room
to prevent undesired contaminants.

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16. 3.9. Harvesting and post harvesting (yield
measures)


Measuring of Parameters such as:
 rate of mycelial extension, number and weight of mature
mushrooms
•pin holes, pileus diameter and stipe length of mushrooms were
recorded
•Mature mushrooms were picked by hand & preserved
•This was done for 4 consecutive flushes (Iqbal et al., 2005).

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17. 4.10. Biological efficiency (yield) measures

Biological efficiency was determined as the ratio of fresh
mushrooms harvested (g) per gram of dry substrates and
expressed as a percentage (Mamiro et al., 2014).

Efficiency (BE) = weight of fresh mushroom
------------------------------ × 100 %
Dry weight of substrate

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18. 3.11. Data analysis

• Data was analyzed statistically on the basis of substrates and
yield parameters per flush on dry weight bases using SPSS
window version.
•Analysis of Variance (ANOVA) was used to indicate
significant mean differences. (Significance level was defined
using (P<0.05).
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19. 4. Results and discussions

4.1.1. Pure cultures
Mycelia invaded full of 10 the PDA plates with diameter of 14 cm
maintained at temperature ( 25°C ),colored cottony within 7 days












P. ostreatus culture on PDA 3-7 days old (a-e ) respectively


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3
4
6
5
7

20. 4.1.2. Comparative study of spawn
production(temperature)

The spawns fully colonized well at room temperature within 14 days but
relatively slower mycelial rate was observed above 30-35°C















25 °C 30°C 35°C 40°C








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21. Comparative study of spawn…
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Effect of Temperature 25-40°C on mycelial growth on spawn medium has
inverse relation ship

22. 4.1.3. Rate of mycelial invasion

•The mother spawn grains in the inoculated substrate just start
colonization after 24h of spawning, until complete invasion of
the whole substrate was seen within the range of 16-35 days.








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CbZ
CbZ*Tfs CbZ*SdC
7days
16 days

23. 4.1.3.Rate of mycelial invasion…

The mean rate of mycelial running among substrate types used
was varied.
The highest running rate was observed in CbZ*SdC1:0.75
(0.70 cm/day) &CbZ (0.69 cm/day), CbZ*Tfs1:0.25
the lowest mycelial running rate Cd*Tfs (0.37 cm/day)
in Cd and in all Cd*SdC treatment bags was observed
(Figur3).


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24. 4.1.3. Mycelial invasion corresponding to
mixtures of substrate treatments bags.

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Cd*SdC
&
Cd*Tfs
Tfs*Cha
(1:0.75)&
Cd control

25. 4.1.4. Pin head initiation.

Temperature, varying due to metabolic reaction from
24ºC to 31ºC and monitored by:
 air exchange (sensitive), prevent fast drying of pin
heads.
In doing so, CO
2 concentration was adjusted by spraying
water .

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Cd *CbZ
CbZ*SdC
CO
2
Cha*CbZ
CbZ*Tfs

26. 4.1.5. Maturation period of oyster fruit bodies
(culture –fruiting)

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Steps of development: pure plate culture, mother oyster spawns ready for use,
primordia, pinheads emerging out 2-3day, fruiting 3days old, mature (5days
old)
CbZ*SdC
CbZ*SdC
CbZ*Tfs

27. 4.1.6.Mean duration of pinning to harvesting
in treatment bags

Only few cases were completed maturation in 6 days after pinning head.
Most treatment bags were harvested within 3-5 days.

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28. 4.1.7. Yield parameters on P.Oysteretus
mushroom

4.1.7.1.Stipe length of substrates

CbZ*SdC
CbZ*Tfs
CbZ
 Cd*Tfs
Cd*CbZ
Cha*SdC&
Control groupswere
moderate
Cd*Cha&Tfs
*Cha
(Relatively shorter
stipe length)
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29. 4.1.7.2. Number of pinning holes of
treatment bags















Effects of Treatment combination on the pinning hole of the mycelium


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30. 4.1.7.3.Pileus diameter


 All of treatment combination which involving corn cobs show
the highest, (Cd*Cb1:0.50,Cb*SadC1:0.25 and Cb control)
and Cd*Tfs1:0.5) were significant different on Pileus diameter
where as Cd control and Cd*SadC were totally unproductive
(Figure 6).

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31. .

4.1.8. Products per flush of substrates


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The results indicated that treatment type did rate the four products
differently.
There was a significant difference across the four flash in mushroom yield,
p<0.001 and
significant differences between treatment groups, p<0.001.
The yield decreases from 1-4
th
flushes.

32. 4.1.9. Biological efficiency of substrates(B.E)
The highest BE among
combination of substrates:
CbZ*Tfs (1:0.75; 1:0.25
and 1:0.50 87.5%, 79.5%
and 72.8%) BE was
observed respectively.
83.6% (CbZ) straw alone.
Followed by the
combination of Cd*Tfs
(1:0.25) with 79.9%
 All treatment CbZ*SdC
(62.6-70%) BE were
analyzed
The lowest yield
(46-50.16%) were obtained
from Cd*ChC and
Tfs*ChC replicants.
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33. 4.1.9.Biological efficiency of substrates…
In general, from this study it may be difficult to conclude, Stipe
length, pilus diameter and other parameters as indicative of
the higher in yield (B.E).
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34. 5. CONCLUSIONS AND RECOMMENDATIONS

5.1. Conclusions
Our results conclude that:
mixtures of all Tfs*CbZ and CbZ*SdC, Cd*Tfs and Chat
(Catha edulis) replicate treatments were the first of its kind and
by its proportion of substrates dosage.
• So used as baseline, practical, and easily accessible alternative
substrates for P. ostreatus cultivation.
•Chat (Catha edulis) steam, shoots discarded as not chewable by
man was suitable substrate for P.ostreatus mushroom cultivation.


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35. 5.1.Conclusions…
•In doing so, reduction of rod side deposited of hazards
environmental pollutant west materials was considered as another
beneficiary aspect.
•In addition CbZ and Tfs (100%) was suitable for the production
of mushrooms with better yield parameters
• All treatment bags promoted mycelial growth, provide yield
parameters and BE except Cd (100%) and all Cd*SdC.




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36. Conclusions
Culture
preparation
(transfer)
Spawn
preparation
Substrates
preparation
Cultivation
(Spawning)
Controlling
environment
al conditions
Management
of pest and
disease
Processing
and
preservation
Aseptic
techniques
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37. 5.2. Recommendations


1. Mushroom cultivation practice should be encouraged by higher institutions,
governments, non governments and all concerned bodies due to;
 agro west materials availability in large quantities and low cost both for
academic trainers as well as sustainably scale up the growth of
Tfs*CbZ and CbZ
 CbZ*SdC,
 Cd*Tfs substrate which were the most effective.
 And chat left over provide moderate yield for commercial scale production to
overcome nutritional, economical and medical related problems in the western
part of the country.

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38. End.






Oct, 2016.