EVALUAT ION OF SOME INVASIVE WEED SPECIES AS A
SUBSTRATE FOR OYSTER MUSHROOM (Pleurotus spp.)
CULTIVATION
MSc Thesis
MINTESNOT BIRARA
April 2012
Haramaya University
EVALUATION OF SOME INVASIVE WEED SPECIES AS A
SUBSTRATE FOR OYSTER MUSHROOM (Pleurotus spp.)
CULTIVATION
A Thesis Submitted to the School of Graduate Studies
(Department of Biology)
HARAMAYA UNIVERSITY
In Partial Fulfillment of the Requirement for the Degree of
MASTER OF SCIENCE IN MICROBIOLOGY
By
Mintesnot Birara
April 2012
Haramaya University
School of Graduate Studies
Haramaya University
As Thesis Research advisors, we here by certify that we have read and evaluated this thesis
prepared, under our guidance, by Mintesnot Birara entitled “Evaluation of some Invasive Weed
Species as a Substrate for Oyster Mushroom (Pleurotus spp.) Cultivation” We recommend that it
be submitted as fulfilling the thesis requirement.
Amare Ayalew (PhD)
Major Advisor
Ameha Kebede (PhD)
Co Advisor
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As members of the Board of Examiners of the MSc Thesis Open Defense Examination, we certify
that we have read, evaluated the thesis prepared by Mintesnot Birara and examined the
candidate. We recommend that the thesis be accepted as fulfilling the thesis requirement for the
Degree of Master of Science the Department of Biology, in Natural and Computational Science.
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DEDICATION
This piece of work is dedicated to my father ATO BIRARA HAILU, and my mother WOIZERO
KIRKIM BAYUH, as well as my sisters.
ii
STATEMENT OF THE AUTHOR
First, I declare that this thesis is my genuine work and that all sources of materials used for the
thesis have been duly acknowledged. This thesis has been submitted in partial fulfillment of
the requirement for MSc degree at Haramaya University, and it is deposited at the University
Library to be made available to users under rules of the library. I solemnly declare that
this thesis is not submitted to any other institution anywhere for the award of any academic
degree, diploma, or certificate.
Brief quotations from this thesis are allowable without special permission provided that
accurate acknowledgement of sources is made. Requests for permission for extended
quotation from or reproduction of this manuscript in whole or in part may be granted by the head
of the major department or the Dean of the School of Graduate Studies when in his or her
judgment the proposed use of the material is in the interests of scholarship. In all other instances,
however, permission must be obtained from the author.
Name: Mintesnot Birara
Signature-----------------------
Place: Haramaya University, Haramaya
Date of Submission: ---------------------------------------
iii
LIST OF ACRONYMS AND ABBREVIATIONS
AOAC
Association of Official Analytical Chemists
ADF
acid detergent fiber
BE
biological efficiency
CF
crude fiber
CP
crude protein
DES
dietary energy supply
FAO
Food and Agriculture Organization
NDF
neutral detergent fiber
PDA
potato dextrose agar
PMC
percentage moisture content
POM
percentage organic matter
SMC
Spent mushroom compost
SMW
Spent mushroom waste
PR
production rate
SR
Spawn running
TA
total ash
USDA
United State Department of Agriculture
WRF
White-rot fungi
TY
total yield
iv
BIOGRAPHICAL SKETCH
The author was born on August 19, 1987 at a place known as Gimbie in West Wollega, Oromia
Region. He completed his junior school at Gimbie Adventist Primary School and high school
education at Akaki Adventist Secondary School. After successfully passing the Ethiopian
School Leaving Certificate Examination (ESLCE), he joined Haramaya University in 2003 and
earned Bachelor of Education Degree in Biology in 2006. After graduation, he has been
working in Addis Ababa as a biology teacher at Akaki L’Esperans Secondary School and School
of Aygoda at a place known as Saris. In June 2010, he joined the School of Graduate Studies of
Haramaya University to pursue his MSc study in Microbiology.
v
ACKNOWLEDGEMENTS
First of all, I would like to praise the Almighty God for his mercy and support in my success,
without His supernatural help nothing is possible. Great Thanks.
I earnestly express my sincere and heartfelt gratitude to my major advisor Dr. Amare Ayalew
and to my co advisor Dr. Ameha Kebede for their valuable technical advice, constructive
comments, and critical review of the thesis with their utmost cooperation and assistance extended
to me during the research write up.
I wish to express my sincere thanks to Dr. Niguse Dechasa and Dr. Mengistu Urge, for providing
me their unfailing assistance during the research and in facilitating all requirements throughout
the period of the study.
I would like to extend my heartfelt appreciation and thanks to Mr. Anteneh Argaw for assisting
me financially and giving me his constructive comments during my research. My gratitude also
goes to staff members of Soil and plant pathology Section, Mr. Yitages, Mr. Girmay, Mrs.
Haymanot, Mrs. Wegayehu, Mrs. Kidist, Mrs. Marta and mushroom project workers Mrs. Rahel,
Mrs. Frehiwot, Mr. Shafi and Mr. Jemal for their invaluable cooperation and during the research
work.
My deepest gratitude goes to my sisters Banchiamlak Birara and my other sisters who offered me
comprehensive moral support and kind treatments that enabled me succeed throughout my study.
vi
TABLE OF CONTENTS
STATEMENT OF THE AUTHOR
iii
LIST OF ACRONYMS AND ABBREVIATIONS
iv
BIOGRAPHICAL SKETCH
v
ACKNOWLEDGEMENTS
vi
LIST OF TABLES
x
LIST OF FIGURES
xi
LIST OF TABLES IN THE APPENDIX
xii
LIST OF FIGURES IN THE APPENDIX
xiii
ABSTRACT
xiv
1. INTRODUCTION
1
2. LITERATURE REVIEW
4
2.1. Mushroom Production and Consumption
4
2.2. Significance of Mushrooms
5
2.2.1. Economic and Environmental importance
5
2.2.2. Biodegradation of organic waste by oyster mushroom and its residual effects
6
2.2.3. Nutritional value of mushrooms
8
2.2.4. Medicinal value of mushrooms
9
2.3. Cultivation of Oyster Mushrooms
10
2.3.1. Substrates used for oyster mushroom production
10
2.3.2. Environmental conditions for cultivation of oyster mushrooms
11
2.3.2.1. Temperature and relative humidity
12
2.3.2.2. Oxygen and carbon dioxide concentration
12
2.3.2.3. pH
13
2.3.2.4. Light
13
2.3.3. Spawning and fruit body formation of oyster mushroom
14
2.3.4. Maturity and Harvesting of Pleurotus spp.
14
2.4. Invasive Weeds as Substrates for Oyster Mushroom Production
15
2.4.1. Potential for use as mushroom substrates
15
2.4.2. Description of target invasive weed species
16
vii
TABLE OF CONTENTS (continued)
2.4.2.1. Parthenium hysterophorus
16
2.4.2.2. Prosopis juliflora
17
2.4.2.3. Lantana camara
17
2.4.3. Chemical compositions of invasive weeds and common substrate for oyster
mushroom
18
2.4.3.1. Wheat straw
18
2.4.3.2. Parthenium hysterophorus
18
2.4.3.3. Prosopis juliflora
19
2.4.3.4. Lantana camara
20
3. MATERIALS AND METHODS
21
3.1. Experimental Site
21
3.2. Experimental Materials and Procedures
21
3.2.1. Source of mushroom strain
21
3.2.2. Spawn preparation
21
3.2.2.1. Preparation and sterilization of spawn substrate
21
3.2.2.2. Inoculation of spawn substrate
22
3.2.2.3. Multiplication of spawn from mother culture
22
3.3. Substrate Preparation and Spawning
23
3.4. Experimental Design
24
3.5. Data Collection
25
3.5.1. Phenological observation
25
3.5.2. Yield related parameters
25
3.5.2.1. Total yield
25
3.5.2.2. Biological efficiency
25
3.5.2.3. Production rate
25
3.5.3. Proximate analysis
26
3.5.3.1. Moisture content (%)
26
3.5.3.2. Crude protein (%)
26
3.5.3.3. Crude fiber (%)
27
3.5.3.4. Total ash and percent organic matter
28
viii
TABLE OF CONTENTS (continued)
3.6. Data Analysis
28
4. RESULTS AND DISCUSSION
29
4.1. Effect of Organic Substrates and Mushroom Species on Growth and Yield
29
4.1.1. Mycelium invasion
29
4.1.2. Pin-head formation
30
4.1.3. Fruiting body formation
31
4.1.4. Yield of oyster mushroom
32
4.1.5. Total yield, biological efficiency and production rate
34
4.2. Effect of Organic Substrates and Mushroom Species on Quality Parameters
37
4.2.1. Percentage moisture content
37
4.2.2. Percentage of dry matter
39
4.2.3. Total ash content
40
4.2.4. Percentage organic matter
41
4.2.5. Crude protein
42
4.2.6. Percentage crude fiber
43
4.3. Correlation Among Parameters
44
4.3.1. Correlation between mushroom yield parameters
44
4.3.2. Correlation between mushroom quality parameters
45
5. SUMMARY AND CONCULUSION
47
5.1. Summary
47
5.2. Conclusions
49
5.3. Recommendation
49
6. REFERENCE
50
7. Appendix
61
7.1. Appendix I. Analysis of Variance Tables
62
7.2. Appendix II Chemical Composition of the Substrates
66
7.3. Appendix III Oyster mushroom cultivation on different substrate
66
7.4. Appendix IV Meteorological Data
69
ix
LIST OF TABLES
Table
page
1.Effect of substrate types on mycelium invasion of Pleurotus species ....................................... 30
2.Effect of substrate types on time taken to pin head formation in Pleurotus species ................. 31
3.Effect of substrate types on fruit body formation in three Pleurotus species ............................ 32
4.Effect of substrate types on total yield (g /2kg) of Pleurotus species ....................................... 35
5.Effect of substrate types on biological efficiency (%) of Pleurotus species ............................. 36
6.Effect of substrate types on production rate of Pleurotus species ............................................. 37
7.The moisture content (%) of Pleurotus species grown on different substrate ........................... 38
8.The percent dry matter content (%) of Pleurotus species grown on different substrate............ 39
9.The total ash content (%) of Pleurotus species grown on different substrate ........................... 40
10.The organic matter content (%) of Pleurotus species grown on different substrate ................ 41
11.The crude protein content (%) of Pleurotus species grown on different substrate .................. 43
12.The percent crude fiber content (%) of Pleurotus species grown on different substrate ......... 44
13.Correlation coefficient for different yield parameters of mushroom ....................................... 45
14.Correlation coefficient for different quality parameters of mushrooms .................................. 46
x
LIST OF FIGURES
Figure
page
1.Structure of lovastatin ............................................................................................................... 10
2.Spawn preparation of oyster mushrooms. .................................................................................. 22
3.Wheat (left) and Lantana camara (right) substrate preparation ................................................ 23
4. Parthenium hysterophorus (left) and Prosopis juliflora (right) substrate preparation. ............ 23
5.Yield of the oyster mushroom (Pleurotus ostreatus) at different flushes .................................. 33
6.Yield of the oyster mushroom (Pleurotus florida) at different flushes .................................... 33
7.Yield of Oyster mushrooms (Pleurotus sajur caju) at different flushes .................................... 34
xi
LIST OF TABLES IN THE APPENDIX
Appendix Table
page
1.Analysis of variance on days of mycelium formation of Pleurotus species .............................. 62
2. Analysis of variance on days of pinhead formation of Pleurotus species ................................ 62
3.Analysis of variance on days of fruit body formation of Pleurotus species .............................. 62
4.Analysis of variance on Biological efficiency of Pleurotus species.......................................... 63
5.Analysis of variance on Production rate of Pleurotus species ................................................... 63
6.Analysis of variance on Total yield of Pleurotus species .......................................................... 63
7.Analysis of variance on Moisture content of Pleurotus species ................................................ 64
8.Analysis of variance on Dry matter of Pleurotus species .......................................................... 64
9.Analysis of variance on Total ash of Pleurotus species ............................................................ 64
10.Analysis of variance on Percent organic matter of Pleurotus species ..................................... 65
11.Analysis of variance on percent Crude protein of Pleurotus species ...................................... 65
12.Analysis of variance on percent crude fiber of Pleurotus species ........................................... 65
13.Carbon and nitrogen content of substrates ............................................................................... 66
xii
LIST OF FIGURES IN THE APPENDIX
Appendix Figure
Page
1. Mycelium invasion of oyster mushroom on wheat straw (left) and on Lantana camara (right)
....................................................................................................................................................... 66
2. Mycelium invasion of oyster mushroom on Parthenium hysterophorus (left) and on Prosopis
juliflora (right) .............................................................................................................................. 67
3. Fruit body formation of oyster mushroom on Wheat straw (left) and on Lantana camara
(right) ............................................................................................................................................ 67
4. Fruit body formation of oyster mushroom on Parthenium hysterophorus (left) and on Prosopis
juliflora (right) .............................................................................................................................. 68
5: Minimum and maximum temperature and relative humidity data during experimental period
....................................................................................................................................................... 69
xiii
EVALUATION OF SOME INVASIVE WEED SPECIES AS A SUBSTRATE
FOR OYSTER MUSHROOM (Pleurotus spp.) CULTIVATION
ABSTRACT
Invasive weeds species with their large biomass production could be suitable candidates for
mushroom production. In view of this, an experiment was conducted to evaluate some invasive
weed species as a substrate for oyster mushroom (Pleurotus species) cultivation 2011/12 at
Mushroom Research, Production and Training Laboratory of Haramaya University. The
experiment was laid out in factorial combination of four substrates (Lantana camara, Prosopis
juliflora, Parthenium hysterophorus and wheat straw as a control) and three edible oyster
mushrooms in a completely randomized design with three replications. Phenological data
including days to onset of mycelium invasion, days to appearance of pin heads and days to
maturation of fruiting bodies of mushrooms were recorded. Yield and yield related parameters
viz. total yield, biological efficiency and production rate of mushroom were also recorded.
Furthermore, quality parameters of mushroom such as moisture content, crude protein, crude
fiber, total ash, organic matter, and dry matter percentage were examined. Pleurotus ostreatus
on Parthenium hysterophorus and Pleurotus ostreatus on wheat straw gave significantly higher
total yield of 1677.45 gm/2kg of dry substrate and 1538.77 gm/2kg of dry substrate, respectively,
while the lowest yield was obtained from Prosopis juliflora in the case of Pleurotus sajur-caju
(555.43 gm/kg of dry substrate). Highest biological efficiency (83.87%) was recorded for
Pleurotus ostreatus grown on dried Parthenium hysterophorus followed by Pleurotus ostreatus on
wheat straw (76.94%). Significantly lower biological efficiency (27.77%) was recorded for the
mushrooms grown on Prosopis juliflora. Higher production rate of 3.13and 3.11 were recorded
for Pleurotus ostreatus grown on Parthenium and wheat straw, respectively. The highest total ash
content (13.90%) was recorded for Pleurotus florida grown on Lantana camara while the lowest
(6.92%) was for Pleurotus sajor–caju grown on the Prosopis substrate. The protein content
(41.48%) of Pleurotus florida grown on Lantana camara was the highest and the crude protein
content (40.51%) of the same species grown on Parthenium hysterophorus ranked second. The
lowest crude protein (30.11%) was for Pleurotus ostreatus grown on wheat straw that was not
significantly much different from other combinations. The fiber content (12.73%) of Pleurotus
sajur caju grown on wheat straw was the highest the second highest crude fiber content (8.87%)
of Pleurotus florida grown on wheat straw. The lowest crude fiber (5.19%) was recorded for
Pleurotus ostreatus on Prosopis. Results of simple linear correlation analysis showed that TY had
a high positive and significant correlation with BE (r = 1.0***) and PR (r = 0.98***). Total
yield had strong negative correlation to phenological parameters such as time taken (days) for
mycelium invasion (r = -0.73***), for pin head formation (r = -0.79***) and also for fruit body
formation (r = -0.77***); in other words BE and PR were also negatively correlated to the
phenological parameters. On the other hand MOC showed positively significant correlation with
CF (r = 0.49**) and negatively significant correlation with DM (r = -1.0***). DM had
negatively and significant correlation with CF (r = -0.49**). A negative and significant
correlation of TA with OM (r = -1.0***) was observed but in case of CP (r = 0.69***) it was
positive significant and OM it was negatively correlated with CP (r = -0.69***). In general,
results showed that with wide C:N ratio organic substrates, the yield and quality of mushrooms
declined. On the other hand, weed residues with narrow C:N ratio, such as Parthenium
xiv
hysterophorus, contributed to the higher yield and improved quality of mushrooms. Considering
the importance of mushrooms as quality food and additional source of income for the rural poor
and marginal farmers of Ethiopia, future studies are justified to evaluate a large number of
easily available organic substrates for growing mushrooms and to standardize the production
techniques under different agro-climatic conditions across the country.
Key words: Invasive weed species, Mushroom Cultivation, Pleurotus spp.
xv
1. INTRODUCTION
Mushrooms are macroscopic fungi with a distinctive spore-bearing fruiting body typically
produced aboveground (on soil) or on a substrate (Stevenson and Lentz, 2007). Most species
of mushrooms belong to the Basidomycota and Ascomycota. Mushrooms produce
lignocellulosic enzymes, which degrade complex organic matter and absorb the soluble
substances (Chang and Miles, 1989). Mushroom cultivation is a major element in
fermentation industry, which involves the bioconversion of cellulose waste into edible
biomass. The cultivation of mushrooms has a great potential for the production of protein rich
quality food and for recycling of cellulosic agro-residues and other wastes (Singh et al.,
1999).
Mushrooms have been used as food for centuries. Although many cultures use mushrooms,
both for their gastronomic importance and their medicinal value, their use as a functional food
has been more notable in oriental cultures, such as China, where the application of
mushrooms for the maintenance of health has its roots thousands of years ago (Sadler, 2003).
Even though mushrooms could supplement the nutritional needs of people for centuries
during the rainy season, which is a period of grain scarcity, the mushroom eating habit among
the majority of Ethiopian population is very poor (Dawit, 1998). Nowadays there is an
increasing understanding that cultivation of mushrooms could play an important role in the
attempt to mitigate malnutrition especially of protein deficiency, in developing countries. The
potential is particularly immense in sub–Saharan Africa, a region with the highest prevalence
of under–nourishment, where one–in–three people are deprived of access to sufficient food
(FAO, 2006).
Out of 38,000 known mushroom varieties, the most popular are Agaricus bisporus (white
button mushroom), Lentinus edodes (Shiitake or Japanese mushroom), Pleurotus species
(Oyster mushroom), Volvarella volvacea (paddy straw mushroom), Flammulna velutipis
(winter mushroom) and Auricularia polytricha (Jew’s ear mushroom) (Singh et al., 1999).
The white button mushroom is the leading cultivated mushroom, constituting 31.8% of the
world mushroom production. However, the share of this mushroom has declined from 56.2%
in 1986 to 31.8% in 1997 (Chang, 1999). The shiitake mushroom, well known by the
Japanese and other Asians, has become the second most cultivated mushroom in the world.
Together with their habit of using them as food, the Asians have a strong tradition to use
shiitake mushrooms medicinally, dating from more than 2000 years ago (Chang, 1996).
Oyster mushroom (Pleurotus spp.) cultivation ranks third after the tremendous increase in
production throughout the world during the last few decades (Chang, 1999; Royse, 2002).
This oyster mushroom accounted for 14.2% of the total world production of edible mushroom
in 1997 with increase from 7.7% in 1986 (Chang, 1999). Pleurotus has been preferred by
mushroom growers because of its flexible temperature and environmental requirements and as
a result it has more cultivated species than any other mushroom (Zadrazil and Dube, 1992).
According to Dawit (1998), in contrast to many countries in South East Asia, mushroom
cultivation is not widespread in Africa. Interest in mushroom cultivation is increasing and
efforts are being made in Africa. The main problem with interested growers in Africa is
unavailability of quality spawn in sufficient quantities, low productivity, and lack of limited
access to the marketing and low level of awareness about mushrooms as food; even though
there is a problem of spawn the oyster mushrooms are the first to be introduced in to
Ethiopian market (Dawit, 1998).
Oyster mushrooms are rather easy to grow at small scale on a wide range of substrates and
different climatic conditions (Kidane, 2006). Dawit (1998) reported 73.8% bioconversion
efficiency for the oyster mushroom grown on cotton seed waste supplemented with 1% wheat
fiber in Ethiopia. Subsequently, researchers urged to gather basic data on different substrates
and mushroom types to promote the development of appropriate technology in the country.
The cultivation of oyster mushroom offers one of the most practicable and economic method
for the bioconversion of agro-lignocellulosic wastes which become problematic for disposal
(Bano et al., 1993; Cohen et al., 2002).
Parthenium is an invasive weed that was first introduced accidentally into Ethiopia in the
1970s. It was first reported from Ethiopia in 1988 at Dire- Dawa and Harerge, Eastern
Ethiopia (Seifu, 1990) and subsequently found near Desse, North-eastern Ethiopia. It is a
noxious weed that causes health hazards on animals and humans. Prosopis juliflora, which
2
was introduced in the late 1970s and early 1980s, is considered a noxious invader weed in
Ethiopia, as well as in Australia, where it has colonized more than 800,000 hectares of arable
land (Duke, 1983). In the Afar region its aggressive growth has lead to a monoculture,
denying native plants water and sunlight, while at the same time denying its nutrients to the
animals that eat its pods or its leaves (Duke, 1983).
Also Lantana camara was implicated in widespread loss of native plant species diversity via
limitation of native species and changed ecosystem structure and function (Gooden et al.,
2009). In this case suitable utilization of weeds is a subject of interest as most weeds are not
used even as fodder due to the presence of lignin and anti-metabolites like phenolics,
glycosides, flavonoids and other compounds (Fianu et al., 1981).The disposal of these plants
through burning causes environmental pollution as they release high level of CO2 as well as
resulting in unnecessary wastage of large amount of organic materials (Croan, 2000). Oyster
mushrooms require carbon, nitrogen and other elements for growth and because of this they
depend on organic compounds as their nutritional sources (Chang, 1999). They can utilize
almost all agricultural wastes as substrates including weed (Miles and Chang, 1997; Taye et
al., 2009). Such plant materials can be used as substrate for primary decomposers such as
white-rot fungi and which have lignocellulosic-degrading enzymes (polyphenol oxidases,
peroxidases etc.) causing significant phenolic removal (Fountoulakis et al., 2002; Aggelis et
al., 2003; Olivieri et al., 2006). Despite the massive research efforts that are underway to
manage these invasive weeds with emphasis on biological control using pathogens and insects
(Taye et al., 2009), immediate and complete control of the weeds could not be expected.
Therefore, an attempt to convert the weed biomass into protein rich food through mushroom
cultivation is prudent.
Thus, this study was undertaken with the following objectives:
 To determine the yield of three species of oyster mushrooms (Pleurotus florida,
Pleurotus sajor caju and Pleurotus ostreatus) cultivated on Lantana camara, Prosopis
juliflora and Parthenium hysterophorus weeds as a substrate.
 To evaluate the effect of the weed substrates on some quality parameters (moisture
content, crude fiber, crude protein, total ash, organic matter) of oyster mushrooms.
3
2. LITERATURE REVIEW
2.1. Mushroom Production and Consumption
The habitual use of mushrooms is well documented in several cultures and religions. They
began to be used as food and medicine in 600 B.C., in Asia. At first, they were harvested in
forests only, and some time later they began to be cultivated by man. The Chinese were
pioneers in the development of fungi culture techniques, shiitake being the first mushroom
produced, by using tree logs (Bernardi et al., 2008). Later, the culture spread to several
countries of North America and Europe (Kues and Liu, 2000). In the 1950s, Asian immigrants
spread cultivation techniques of edible mushrooms from their original countries.
In the last few years, worldwide mushroom production has increased over 300%, reaching
approximately 2.96 metric tons in 2002 (USDA, 2003). China has become the top-producing
nation for all edible mushrooms, turning out over 40% of the world's supply (USDA, 2003).
The U.S. is the next largest producer of mushrooms, contributing about 13%, while the
Netherlands and France produce about 9.5 and 5%, respectively (USDA, 2003). Overall
U.S. production by volume has been steadily rising over the last decade. Operations are also
diversifying, adding production of various specialty mushrooms. Industry expansion, in both
output and diversity, is largely due to improvements in cultivation technologies and the
expansion of market demand (Yamanaka, 1997).
Although there are more than 2000 species of edible mushrooms nowadays, the button
mushroom (Agaricus bisporus), the oyster mushrooms (Pleurotus ostreatus and Pleurotus
ostreatoroseus) and shiitake (Lentinula edodes) are among the most cultivated ones (Bononi
et al., 1999). Oyster mushroom cultivation can play an important role in managing organic
wastes whose disposal has become a problem (Das and Mukherjee, 2007).
According to Dawit (1998), in indigenous forests, edible mushroom species of Macrolepiota,
Auricularia, Armillaria, Pholiota, occur in abundance and several species of Macrolepiota
and Agaricus are common on highland grazing areas in south west. Mushrooms associated
4
with termites, Termitomyces species, are diverse and nutritious. Several edible mycorrhizal
mushrooms are associated with exotic trees such as Eucalyptus, Cupressus and Pinus. The
need to identify edible from poisonous mushrooms is very important in the promotion of
mushroom foods. In Ethiopia, the habit of mushroom eating differs from region to region and
among the different ethnic groups. Mushroom eating habit of people in the south and
southwest is more developed than people on the central and northern highlands of Ethiopia
who are not accustomed to mushroom foods consumption (Dawit, 1998).
Though mushrooms could supplement the nutritional needs of people during the rainy season
which is a period of grain scarcity, the mushroom eating habit is very poor in certain regions.
Contrary to this, the Majangir people, (in gambella national regional states), for instance,
have a well developed habit of mushroom utilization. Mushrooms are highly valued and
consumed during the rainy season. Mushroom growing is a new economic activity in Ethiopia
and the three most important cultivated mushrooms, Agaricus bisporus (button mushroom),
Pleurotus ostreatus (oyster mushroom) and Lentinula edodes (shiitake mushroom) are
commercially grown on small scale and supplied to hotels and supermarkets in Addis Ababa
(Dawit, 1998).
2.2. Significance of Mushrooms
2.2.1. Economic and Environmental importance
Pleurotus ostreatus is a prospective source of valuable food protein, and an organism with the
ability to utilize various lignocellulosic materials (Wang et al., 2001). In addition, the
substrate, used for mushroom cultivation, is valuable as fertilizer and soil conditioner for the
growth of plants, following the harvesting of the mushrooms (Brenneman and Guttman,
1994). Additionally, fermented residues could be used as animal feed after mushroom
cultivation (Soto-Cruz et al., 1999). The technology can also limit air pollution associated
with burning agriculture wastes as well as to decrease deleterious fungal inoculums
populations the utilization of waste paper for the production of cultured mushroom can solve
one of the most important problems in solid waste disposal. This will provide an economical
5
gain and protect the environment while providing a nutritious food source from mushrooms
(Pandey et al., 2000).
Agriculture is the mainstay of Ethiopian economy. Due to the growing population and high
demand for land, producing much on small plot of land is a major issue. Mushroom
cultivation is a space-confined technology and requires relatively small capital. With
declining land productivity and increasing interest in organic farming, the cultivation of edible
saprophytic mushrooms offers prospects for using agricultural residues (Shasho, 2004).
Cultivation of edible mushrooms is one of the most economically viable processes for the
bioconversion of lignocellulosic wastes (Bano et al., 1993; Cohen et al., 2002).Various
agricultural by-products are being used as substrates for the cultivation of the oyster
mushroom. Some of these wastes include banana leaves, peanut hull and corn leaves, mango
fruits and seeds, sugarcane leaves, wheat and rice straw (Cangy and Peerally, 1995).
Approximately 100,000 tons of coffee pulps are generated each year in Mexico, and the
majority of this waste has no further economic use; instead, coffee growers generally spread it
in the field where it is allowed to decompose. Diverse technologies have been proposed for
utilizing the byproducts generated by the coffee industry (Pandey et al., 2000). Culturing
edible mushrooms on coffee pulp seems especially attractive, since it represents a direct
conversion of an agricultural waste to human food. Among edible mushrooms evaluated for
this commercial activity, Pleurotus strains appear promising, primarily because their
biological efficiencies can exceed 100% (wet base) (Martı´nez-Carrera et al., 1985;
Martı´nez- Carrera, 1989).
2.2.2. Biodegradation of organic waste by oyster mushroom and its residual effects
The biodegradation of lignocellulosic by-products from agriculture or forestry confers
ecological importance on mushroom cultivation (Wood, 1985). Laboratory studies on the
degradation of lignocellulose, including wood, straw, and cereal grains, have focused mainly
on a few fungal species that grow well in the laboratory and can be readily manipulated in
liquid culture to express enzymes of academic interest. The white-rot fungus (WRF)
6
Pleurotus ostreatus is an edible basidiomycete known for its ability to degrade agro-industrial
lignocellulosic wastes, which are mainly composed by cellulose, hemicelluloses, and lignin. It
is generally cultivated on wheat straw, but other lignocellulosic substrates, such as cotton
stalks, have proved adequate for its growth (Yildiz et al., 2002). Bioconversion of
lignocellulosic residues through cultivation of Pleurotus species offers the opportunity to
utilize renewable resources in the production of edible, protein-rich food that will sustain food
security for people in developing countries (Sanchez et al., 2002).
Several WRF (P. ostreatus, P. pulmonarius, Lentinula edodes and Hypsizygus tessellates)
have been cultivated for mushroom production. In mushroom production, for example, 5 kg
of spent (used up) mushroom waste (SMW) will be generated from the production of every
kilogram of mushrooms (Semple et al., 2001). High levels of residual nutrients and enzymes
are still present in SMW. When using WRF for bioremediation, the availability of fungal
inocula is a practical consideration and the use of SMW could be advantageous due to the low
cost and environmentally-friendly treatment.
Since more than 25% of world wide mushroom production is Pleurotus mushrooms, it would
be advantageous to use Pleurotus SMW. Yet improvement is needed for shortening the time
and reducing the price. The spent mushroom composts (SMCs) of Agaricus and Pleurotus as
wastes of mushroom industry, still contain many residual enzymes, e.g. proteases, cellulases,
hemicellulases, lignin peroxidase, manganese peroxidase and laccase (Lau et al., 2003). The
last three enzymes belonging to lignolytic enzymes act as Fenton reagents to produce reactive
radicals for non-specific cleavage of a wide variety of highly recalcitrant organopollutants
(Gong et al., 2006).
White-rot fungi appear to have a great potential in the degradation of a wide range of organic
substrate. This capability is based on the production of extracellular peroxidases (manganese
and lignin peroxidase and laccases) that catalyze the breakdown of organic compounds (Gadd,
2001; Hattaka, 1994). Most studies concerning organic compounds degradation have been
done using Phanerochaete chrysosporium. However, there has been an interest in screening
new species, which can produce higher levels of ligninolytic enzymes and a greater ability in
degrading pollutants. Pleurotus ostreatus has been tested for its ability to degrade lindane
7
(pesticide) under different conditions (Sheeja and Murugesan, 2002). It is the first time that
Pl. ostreatus has been used in lindane degradation, although other species of Pleurotus such as
florida, sajor-caju, eryngii and different white-rot fungi have been found to degrade this
pesticide (Tekere et al., 2001).
2.2.3. Nutritional value of mushrooms
In general, mushrooms are quite high in protein, with an important content of essential amino
acid, but low in fat (Mattilda et al., 2001). Furthermore, these fungi supply a large amount of
carbohydrates and fiber and a nutritionally significant content of vitamins (B1, B2, B12, C
and D) and mineral elements (Ca, K, Mg, Na, P, Cu, Fe, and Mn) (Mattilda et al., 2001).
Mushrooms have been used as a food for centuries. The mushroom shiitake, well-known by
the Japanese and other Asians, has become the second most cultivated mushroom in the
world. Together with their habit of using them as food, the Asians have a strong tradition to
use mushrooms medicinally, dating from more than 2000 years ago (Chang, 1996).
As Yang et al., (2001) reported crude protein content on dry weight basis is 15.4% and 23.9%
in P. cystidiosus and P. ostreatus, respectively. They contain about 60% carbohydrates (dry
weight), within the ranges for other edible mushrooms (Crisan and Sands 1978). In addition,
they were reported to be low in fat (2 to 3% by dry weight), a good source of essential amino
acids, and contain approximately 5 to 9% fiber (Yang et al., 2001). Research into mushrooms
and their potential role in weight loss diets is currently underway. One 80 g serving provides
only 10 kcals and 0.4g of fat. In addition, mushrooms’ high water content (about 90%) can
contribute to a feeling of fullness, and low energy (calorie) density can help to promote
weight maintenance. Chitosans are used in slimming products, as they lower the absorption of
lipids from food. Their capacity to reduce body weight has been demonstrated in studies
performed on two groups of volunteers, each comprising 100 people on a low energy diet. In
participants who had been administered two chitosan tablets for four weeks, a loss of over 7
kg surplus weight loss was observed, while in the control group the average weight loss was 3
kg (Muzzarelli, 1999).
8
2.2.4. Medicinal value of mushrooms
For centuries, mushrooms have been prescribed for various ailments. Research has shown that
some of these claims are not mere tradition but having some scientific basis. A vast body of
information exists in the scientific literature, dating back to the 1940’s and 1950’s (Quimio et
al., 1990). Studies in the period of the 1960’s in Japan and the United States, have shown that
cultivated mushrooms such as Agaricus bisporus, Lentinula edodes and Pleurotus species
contain a high amount of retene a substance that may, in some circumstances, have an
antagonistic effect on some forms of tumor (Binding, 1972).
Traditionally, mushrooms are highly valued as folk medicines and functional foods for their
antitumor and other physiological benefits. Ganoderma was found to be medically active in
several therapeutic effects including anti inflammatory, antitumor, antiviral (e.g. anti-HIV),
antibacterial and antiparasitic, blood pressure regulation, cardiovascular disorders,
immunomodulating, kidney tonic, hepatoprotective, nerve tonic, sexual potentiator and
chronic bronchitis (Wasser and Weis, 1999).
Oyster mushrooms are also known to have multiple medicinal properties. Two of the more
prominent medical attributes are cardiovascular and cholesterol-controlling benefits. Oyster
mushrooms naturally produce mevinolin (lovastatin) (Figure 1) in portions of the fruiting
bodies (Gunde-Cimerman, 1999). Mevinolin inhibits the key enzyme in cholesterol
biosynthesis in the liver and reduces cholesterol absorption (Bobek et al., 1998). P. ostreatus
is a known producer of many biologically active substances. It has been demonstrated to have
antibacterial properties (Wasser and Weis, 1999) in addition to antiviral, anti-inflammatory
and immune modulation activities (Jose et al., 2002). It is also believed to be effective in the
treatment of cancer. Gunde-Cimerman (1999) showed its effectiveness as an anticancer agent,
while Gerasimenya et al. (2002) found it useful in decreasing the toxic effects of common
cancer drugs.
9
Figure 1 : Structure of lovastatin Source: http:// bioweb.uwlax.edu
2.3. Cultivation of Oyster Mushrooms
2.3.1. Substrates used for oyster mushroom production
Oyster mushroom can be cultivated in any type of ligno cellulose material like straw, sawdust,
rice hull, etc. The choice of substrate will depend on the availability of suitable agricultural
waste in a particular country (Quimio et al., 1990). In Ethiopia, since 85% of the people live
in rural sectors, it is obvious that there are alternatives, such as biomass of weeds. But
according to the literature slight work has been reported on nutritional component and
substrate usage of the plant conserning these materials for growing mushrooms (Fianu et al.,
1981).
Hami (1990) studied the oyster mushroom cultivation on sawdust of different woods and
found that P.ostreatus gave the maximum yield. Presently sawdust is commonly used and is
the preferred medium at commercial scale. Hami (1990) reported that P.ostreatus gave
maximum biological efficiency among sawdust from different trees. Of the sawdust types,
softwood sawdust like mango and cashew are known to be more suitable than hardwood
sawdust. Dawit ( 1998) described that oyster mushrooms could be cultivated on a wide range
of lignocellulosic materials such as tef, bean pod, wheat, rice, maize, sorghum, cotton waste,
cotton seed waste, coffee seed waste or pulp, synthetic waste paper, banana peels, corn cobs,
bagasse, and palm pericarp waste.
10
According to Stamets (2000), Pleurotus species, with a 24.2% of world production, have the
ability to grow directly on unfermented agricultural wastes. Mushroom cultivation is a
worldwide practice which utilizes almost all agricultural and agro-industrial residues as
substrate (Chang, 1999). Various techniques are used to prepare substrate for the cultivation
of Pleurotus mushrooms (Villa-Cruz et al., 1999; Geml et al., 2001). One of these is
composting, which, when carried out properly, reduces the level of competitive
microorganisms in the material being prepared. Microorganisms indigenous to the byproducts
metabolize other compounds, including sugars and generate heat during metabolism.
As a result, the substrate reaches temperatures ranging from 50 to 70o C for several days
(Stamets and Chilton, 1983; Laborde et al., 1993). Composting has been successfully applied
in the preparation of a substrate selective for Pleurotus spp. with biological efficiencies of
70% achieved in three harvests (Villa-Cruz et al., 1999). According to Quimio et al (1990),
Pleurotus species can be grown on various agricultural waste materials with the use of
different technologies. They grow well on different type of lignocellulosic materials,
converting the materials in to digestible and protein rich substances suitable for animal feeds.
Oyster mushroom is a white-rot fungus that uses lignin and cellulose together as its carbon
source (Kang, 2004). The C: N ratio is important factor for optimal substrate composition for
mushroom. Therefore, any type of organic matter containing lignin and cellulose can be used
as oyster mushroom substrates, and this includes almost all agricultural wastes.
2.3.2. Environmental conditions for cultivation of oyster mushrooms
When choosing or creating a site for mushroom production, consideration of environmental
conditions is important. Environmental factors include temperature, relative humidity, light,
carbon dioxide and acidity of substrate, which vary together in their co–dependent
relationships. As the growing room temperature is raised, relative humidity decreases. A
higher temperature promotes fruit body metabolism, which in turn, increases their respiration
rate and results in high carbon dioxide production. Oyster mushroom needs different
environmental condition at each growing stage. During incubation, appropriate relative
humidity of 65–70% and water content of 65% substrate is required (Kang, 2004). Optimum
11
temperature for mycelium growth is 20–25oC. Pleurotus florida reaches its optimum growth
at 25oC but some strains reach optimal growth at 25–35oC, which suggests that they are a
good choice for cultivation in both temperate and tropical regions (Kang, 2004).
2.3.2.1. Temperature and relative humidity
Culture of oyster mushroom is becoming popular throughout the world because of their
abilities to grow at a wide range of temperatures and to utilize various lignocelluloses (Baysal
et al., 2003). Oyster mushrooms are able to grow and thrive in a wide range of temperature
environments. Stamets (2000) recommends temperatures between 10°C and 21°C for
development of oyster mushrooms. Pettipher (1987) achieved successful fruiting of
P.ostreatus with daily temperatures ranging between 8°C and 33°C. Extremely high humidity
(90 to100%) is recommended for optimal primordial formation. Once primordia have formed,
humidity should be lowered to 85 to 90%. Ideally, humidity levels should be managed so that
mushrooms are regularly receiving moisture but excess moisture can evaporate from fruit
body surfaces (Stamets, 2000). Excessive moisture can cause lack of oxygen in the substrate,
as well as encourage certain contaminates. Insufficient moisture can prevent primordia
formation and stunt fruit body growth.
2.3.2.2. Oxygen and carbon dioxide concentration
Since growth of the fungus produces carbon dioxide as it decomposes the substrate,
introduction of 'outside' air reduces carbon dioxide build up and increases oxygen levels.
Fungal mycelium is extremely tolerant of carbon dioxide, thriving at 20% CO2 levels. Oxygen
is required for formation of fruit bodies. A significant decrease in ambient CO2 level and
increase in oxygen is critical for the initiation and development of primordia. Thus sufficient
air circulation within a mushroom fruiting site is vital. Excessive influx of outside air,
however, greatly affects both temperature and humidity of the environment (Stamets, 2000).
In order to test the activities of various mycelium mass, 5% and 10% inoculation material,
were introduced to the substrate and three days after inoculation the carbon dioxide
concentration had already raised to over 20% (Zadrazil, 1975). As expected, the case with
12
10% spawn showed a greater concentration than the case with 5% inoculation. Four and six
days after inoculation, carbon dioxide concentration had reached maximum then decreased,
finally showing no material alteration from the 16th to the 31st day. Carbon dioxide production
is nevertheless dependant on the ingredients in the substrates. The high CO2 concentration in
the substrate serves as a shield for the Pleurotus against other microorganisms, which either
grow or die off at higher concentration (Zadrazil, 1975).
2.3.2.3. pH
Mycelial growth (mycelium weight) was determined in submerged culture with pH values
between 4 and 7 at intervals of 0.5pH by Chang and Hayes (1998). They reported that a very
acid nutrient (pH 4) inhibits the growth of Pleurotus florida and Pleurotus eryngii. Rising pH
values from 4 to 6 affected mycelia growth of both species favorably. The pH optimum for
Pleurotus florida lies between 5.5 and 6.5, while for Pleurotus eryngii the pH is slightly 4
lower, between 5 and 6 with rising pH values, above the optimum, growth is inhibited. The
pH value of the nutrient (solid and liquid) is changed by mycelia growth. In very acid nutrient
medium, the pH value change very slowly since the rate of growth is very slow. With rising
mycelia growth, the pH value changes more rapidly. Within the optimum limits the value
varies only slightly. High pH values (over 6) are lowered by the growth of the mycelium.
2.3.2.4. Light
As a forest–dwelling mushroom, indirect natural light is considered ideal for the formation of
Pleurotus species fruit bodies. Although the mycelium of the oyster mushroom does not
require light, proper fruit body formation requires moderate light. Too little or too much light
can lead to discolored, malformed fruit bodies or the inability to fruit. Kalberer (1974) found
that oyster mushroom yield was maximized using light levels of 60 to 86 μ mol/m 2/sec (300
to 430 lux) for twelve hours per day. Stamets (2000) recommends levels around 200 to 300 μ
mol/m2/sec (1000 to 1500 lux) for commercial production.
13
2.3.3. Spawning and fruit body formation of oyster mushroom
Shah et al. (2004) reported that spawn running takes 2-3 weeks after inoculation. Pin-head
formation is the second stage of mycelial growth during cultivation of mushroom. Small pinhead like structures were observed, these pinheads were formed 6-7 days after spawn running.
Similarly, Ahmad (1986) reported that P. ostreatus completed spawn running with 17-20 days
on different substrates and time for pinhead formation was noted between 23-27 days after
inoculation. Fruiting body formation is third and final stage during the cultivation of oyster
mushroom. The fruiting bodies appeared 3-6 days after pin-head formation and took 27-34
days later after inoculation of spawn (Shah et al., 2004).
According to Jandaik and Goyal (1995) spawn is mixed on wet straw at the rate of 2% and it
is filled in polyethyline bag 3-6 kg/bag. At this rate, Tripathi (2005) reported that one bottle
(270-300g) of spawn is sufficient to seed 3kg dry substrate or 14-15 kg wet substrate. Davis
and Aegerter (2006) suggested that the cleanliness of spawn is absolutely critical and its rate
should be 2.5%. Regarding the age of spawn, the author further stated that freshly prepared
(20-30 days old) grain spawn is best for spawning and spawn of more than one month should
be stored at room temperature to retain its viability. Old spawn usually form very thick mat
like structure due to mycelium aggregation and sometimes young pin-heads and fruiting
bodies start developing in the spawn bottle itself and such spawn should not be used for
spawning.
2.3.4. Maturity and Harvesting of Pleurotus spp.
Jandaik and Goyal (1995) described that the oyster mushroom fruit bodies are picked when
edges of pilei start to fold or curl upward. In contrary Nicola (2003) stated that the right time
of harvesting is when the cap has opened up and the gills of the mushroom are visible but the
edge of the cap is still slightly rolled inwards. However, the harvesting standards for the
oyster mushrooms are different for different products. Picking of the mushroom is done by
twisting the mushroom gently, so that it is pulled out without leaving any stub and disturbing
the fruit bodies. The base of the stipe found within the straw should be removed by cutting off
14
with a sharp knife, because it will cover the new emerging pin part of the mushroom. The
caps of Pleurotus species, to be canned or preserved in brine, should be 4 to 5cm.
Mushrooms should be picked when young, and preferably in cluster. Once the gills produce
abundant spores, storability rapidly declines. Mushroom surfaces should be slightly dry at
harvest (Stamets, 2000). Royse et al. (2004) explained that, mushrooms are harvested from
the substrate the same time each day when the in-rolled margins of the basidiomes began to
flatten. The substrates clinging to the stipe should be cut away.
Oei (2003) states about harvesting time and advises to pick when the caps have almost
completely spread out, about five to seven days from tiny pinhead to the harvested oyster
mushroom. Harvesting begins when the cap is at its maximum size and before the veil has
stretched and opened, exposing the gills. A slight twist of the fingers at the stem where it joins
the substrate will make mushroom come free. Care is required to insure that the cutting of the
attached mycelium does not damage surrounding pins. Mushrooms are harvested when the
cap is from 2.5 to 7.5 cm or large in diameter than the base stem. Since easily damaged, they
must be handled with great care (Libnernonnecke, 1989).
2.4. Invasive Weeds as Substrates for Oyster Mushroom Production
2.4.1. Potential for use as mushroom substrates
Various kinds of weeds have been used as substrates for P. ostreatus cultivation. The
following species were used: Leonotis sp. (Lamiaceae), Sida acuta (Malvaceae), Parthenium
argentatum
(Asteraceae),
Ageratum
conyzoides
(Asteraceae),
Cassia
sophera
(Caesalpiniaceae), Tephrosia purpurea (Papilionaceae) and Lantana camara (Verbenaceae)
(Fianu et al., 1981). These plants were completely sun-dried, sectioned into small pieces and
soaked in water, and, subsequently, excess water was drained. Each species was inoculated
with and without supplementing with wheat straw and afterwards subjected to incubation.
Leonotis sp. supplemented with wheat straw significantly increased P. ostreatus yield
15
(1.30kg-1). It was concluded that the use of weeds as substrates is an efficient method for the
cultivation of edible mushrooms (Fianu et al., 1981).
2.4.2. Description of target invasive weed species
Invasive Alien Species (IAS) refer to plants, animals or micro organisms that are not native to
specific ecosystem and whose introduction threatens biodiversity, food security, health or
economic development (McNeely et al. 2001). Invasive species are of concern because of
their capability of spreading fast, their high competitiveness and ability to colonize new areas
within short periods. The nature and severity of the impacts of these species on society,
economic life, health and national heritage are of global concern (McNeely et al. 2001).
2.4.2.1. Parthenium hysterophorus
Parthenium hysterophorus L., a native subtropical species of North and South America, is
spreading rapidly in many parts of the world (Javaid and Anjum, 2005) Parthenium
hysterophorus commonly known as congress weed, is a flowering plant belonging to the
family Asteraceae. Being a declared invasive weed, it is currently threatening the biodiversity
and human health in several areas of the world. Several researches have documented the
allelopathic effect of this weed. Therefore, parthenium management would remain a great
concern of the century. However, several studies proposed that parthenium can be used as a
green manure, compost, biocontrol, soil ameliorate that may improve physical, chemical and
biological properties of the soils and is a source of readily available plant micro-and macronutrients. Numerous studies revealed that the integrated use of parthenium in soil modifies the
physico -chemical, biological and nutritional quality of the soil. Parthenium has great
potentiality in agriculture due to its efficacy in modification of soil health and crop
performance (Channappagoudar et al., 2007).
The high concentration of elements (N, P, K, Fe, Mn, Cu and Zn) in composted parthenium
increases the yield of many agricultural crops. An exhaustive review of numerous studies of
last two decades took place in the study, which systematically covers the importance, scope
and apprehension regarding utilization of parthenium in agriculture. Parthenium
hysterophorus can be used as a bioherbicide. Appreciable quantity of nutrients in parthenium
16
can be utilized to nourish the crops after composting and a lot of green parthenium can be
destroyed. This suggests that composting of uprooted parthenium, or use as a green manure
and parthenium extract may reduce its spreading and inhibit the weed growth as well as
menace of human health hazards worldwide (Prem Kishor et al., 2010).
2.4.2.2. Prosopis juliflora
In Mexico, Argentina, and Brazil, Prosopis pods are an important source of animal feed
(Felker and Moss, 1996). In Peru, pods of especially sweet varieties are used for human food.
Prosopis juliflora pods are a valuable low cost fodder in the semi-arid areas of northeastern
Brazil (de Barros et al., 1988), where it partly offsets fodder scarcity during the dry season.
Prosopis juliflora, a perennial deciduous thorny shrub, the common vegetation of semi-arid
region of Indian subcontinent, was used as a raw material for the production of cellulosic
ethanol. The leguminous plant of the Prosopis genus found in northern Mexico (mesquite) has
recently been suggested to be used as raw material for long-term sustainable production of
cellulosic ethanol (Hopkins, 2007). Its nature to tolerate drought, grazing, heavy soil, sand as
well as saline dry flats and no competence with animal feed demand made it a potential low
value substrate for ethanol production. Here, an attempt was made to scarify P. juliflora into
reducing sugars and eventually to ethanol fermentation.
2.4.2.3. Lantana camara
Lantana camara L. is a species of flowering plant in the family Verbenaceae grows
abundantly in many parts of the world, and has encroached upon a large expanse of pastures
and wastelands (Sharma et al., 1988). Efforts are being made to look for alternate substrates
for mushroom cultivation involving little competition with the animal feed systems (Hartley,
1987). The plant causes toxicosis in domestic animals and human beings (Sharma et al.,
1991). The cultivation of edible fungi appears economically to be the most viable means of
utilizing lignocellulosic wastes (Bano et al., 1993; Cohen et al., 2002). Lantana substrate has
been used in conjunction with waste paper for cultivation of Agaricus bisporus and Pleurotus
ostreatus (Bist and Harsh, 1983, 1985).
17
2.4.3. Chemical compositions of invasive weeds and common substrate for oyster
mushroom
2.4.3.1. Wheat straw
The study of chemical composition of substrates is very significant because the nutritional
quality is highly correlated to substrate composition (Oei, 2003). Srinivas and Gupta (1997)
reported that wheat straw contains 87.9% organic matter (OM), 0.6% nitrogen, 0.8% ether
extract, 82.2% neutral detergent fiber (NDF), 54.7 % acid detergent fiber (ADF) and 12.1%
total ash. According to Vats et al. (1994), wheat straw contains 0.82% nitrogen, 46.05%
cellulose, 26.61% hemicellulose and 5.95% lignin. A nitrogen content of 0.4% and a C: N
ratio of 80–127 for wheat is reported by Steven (2004).
Adamovic et al. (1998) reported the changes on wheat straw after seeding and showed that
NDF decreased from 824 to 485 g kg-1 and ADF from 561 to 412 g kg-1.They noted similar
tendency for hemicelluloses and cellulose, while it was not so pronounced for lignin. Ash
content increased from 62.6 to 97.8 g/kg. The authors calculated degradation rates for
individual constituent of cell wall components and found hemi cellulose to be 0.902%, lignin
0.450%, NDF 0.402%, cellulose 0.290% and ADF 0.276% per day.
2.4.3.2. Parthenium hysterophorus
Experiments were conducted in India on Parthenium as compost to know some of its
chemical composition and it is reported to contain high amounts of N, P and K (Son, 1995;
Biradar et al., 2006). Channappagoudar et al. (2007) reported that parthenium composted preflowering has higher N content (2.95%) compared to poultry manure (2.02%), vermi compost
(1.21%) and farm yard manure (0.54%). The same study indicated that Parthenium compost
has higher P content (0.82%) compared to farm yard manure (0.26%) but lower as compared
to poultry manure (1.6%) and vermi compost (0.86%). Its K content is also higher (1.39%)
compared to Vermi compost (0.55%) and farm yard manure (0.34%) whereas lower compared
to poultry manure (1.42%).
18
2.4.3.3. Prosopis juliflora
Prosopis juliflora is a leguminous tree that is native to arid and semi-arid regions of North
America (Pasieczink, 2001; Harris et al., 2003). It is present in North America, Africa and
Asia, having green-brown twisted stem, flexible branches, and produce’s flattened, multiseeded curved pods with hardened pericarp (Habit and Saavedra, 1988). The crude protein
and energy contents of P. juliflora pods are comparable to those in barley grain (Abdullah and
Abddel hafes, 2004). Previous studies have shown that replacing wheat bran or barley grains
and corn by improved dry matter intake, average daily gain, and feed efficiency (Abdullah
and Abddel hafes, 2004).
Regarding Prosopis nutritional composition of dried pods, Vimal and Tyagi (1986) have
reported that the pods contain the following nutrients: protein, 16.5%; fat, 4.2%;
carbohydrate, 57%; fiber, 16.8%; ash, 5.4%; calcium, 0.33%; and phosphorus, 0.44%.
Moreover, Gujarat Agricultural University, determined the trace-element composition of
Prosopis pods as 12.46 to 15.51 ppm copper, 22.11 to 22.30 ppm manganese, 18.30 to 28.01
ppm zinc, and 203 to 638.8 ppm iron (Shukla, et al., 1984). Because of the high carbohydrate
content and good amount of protein, the spongy walls of ripe pods are highly nutritive and
used in making meal for humans (Pinole) and alcoholic beverages (mesquite wines, etc.). The
husk of pods is used for dying; they contain tannin (1.9%). Shukla, et al., (1984) have noticed
that the ripe pods are said to have high nutritive value, i.e., rich in sugar and nitrogen and are
greedily eaten by most of the herbivorous animals and livestock. Further, the pods may yield a
substitute for wood shavings used in various industries for thermal insulation and sound
control (Shukla, et al., 1984). Studies on palatability and nutritive value of pods and their
source as livestock feed and milk production, particularly goats, sheep, and camels, have been
conducted in different countries.
The Prosopis flower per 100 g is reported to contain ( dry weight): 21.0 g protein, 3.2 g fat,
65.8 g total carbohydrate, 15.5 g fiber, 10.0 g ash, 1,310 mg Ca, and 400 mg P. Leaves
contain 19.0 g protein, 2.9 g fat, 69.6 g total carbohydrate, total carbohydrate, 21.6 g fiber, 8.5
g ash, 2,080 mg Ca, and 220 g P. Fruits contain 13.9 g protein, 3.0 g fat, 78.3 g total
19
carbohydrate, 27.7 g fiber, and 4.8 g ash. Seeds contain (dry weight) 65.2 g protein, 7.8 g fat,
21.8 g total carbohydrate, 2.8 g fiber, and 5.2 g ash (FAO, 1980).
Another analysis of the fruit shows 14.35% water (hygroscopic), 1.64% oil, 16.36% starch,
30.25% glucose, 0.85% nitrogenous material, 5.81% tannin-like material, 3.5% mineral salts,
and 27.24% cellulose. Mesquite gum readily hydrolyses with dilute sulfuric acid to yield Larabinose and D-galactose and 4-o-methyl-D-glucuronic acid at 4:2:1. Owing to the high
content of arabinose, the gum is an excellent source of sugar. Roots contain 6.7% tannin, bark
3–8.4%, and dry wood 0.9%. The alkaloids 5-hydroxytryptamine and tryptamine are reported
from this species (Simpson, 1977).
2.4.3.4. Lantana camara
Lantana contain appreciable amount of sugar, tannin, resin (a crystalline glucoside, C 27 H44
O4), and several enzymes, such as catalase, amylase, invertase, lipase, tannase and
glucosidase, have been found. The important inorganic elements are: N,1.51%; P(P2O5),
0.15%; K(K2O), 0.90%; Ca (Ca O), 0.61%; Mn, 0.3-0.4%; and ash (rich in minerals), 10.29%.
An idea of the chemical composition of the plant has been given by Nigam et al., (1977).
Another chemical analysis was carried out on lantana (Lantana camara). The proximate
analysis (on dry weight basis) of lantana seeds showed the following composition: moisture
17.27%, and ash 1.81%, fat 11.0% crude protein 6.3%, and carbohydrate 80.9%. Concerning
its minerals composition, potassium was the most abundant element, followed by phosphorus
and sodium. Zinc, iron, copper, and manganese were also detected in it. Analysis of amino
acids revealed that the first limiting amino acid was valine (Hassan and Mokhtar, 2011).
The plant contains appreciable amounts of sugar, tannin, resin (a
20
3. MATERIALS AND METHODS
3.1. Experimental Site
The study was conducted at Haramaya University from January 2011 to April 2012. The
experimental site was at the Mushroom Research, Production and Training Laboratory of
Haramaya University located within the campus at 42°3lE longitude, 9°26lN latitude and at
altitude of 1980 m.a.s.l. (AUA, 1996).The experimental room average minimum and
maximum temperatures were 12.96oC and 25.72oC respectively. Relative humidity was in the
range of 57.67-91.93% have been managed by an instrument known as data loger
(Hygrometer)
3.2. Experimental Materials and Procedures
3.2.1. Source of mushroom strain
Pure cultures of three edible oyster mushroom species (Pleurotus florida, Pleurotus sajor–
caju and Pleurotus ostreatus) were obtained from Mushroom Research, Production and
Training Laboratory of Haramaya University. The pure cultures were initially multiplied on
sterilized potato dextrose agar (PDA) medium to get enough stock culture of the pure species
needed to prepare the required quantity of spawn (Sara, 2007). Stock culture was grown by
transferring the cultures into flasks containing grain (maize) as described under section 3.2.2.1
3.2.2. Spawn preparation
3.2.2.1. Preparation and sterilization of spawn substrate
Mother spawn was prepared according to the procedure described by Singh and Chaube
(1995). Nine kg of the maize seeds were boiled in 15 liters of water for 15 minutes and then
allowed to remain soaked in the hot water for another 25 minutes. The water was drained off
and the grain was put in a sieve to dry over night. Next day, 120 g calcium sulfate and 30 g
calcium carbonate were mixed with each 9 kg of the boiled grains. The calcium sulfate and
21
calcium carbonate were used to maintain the pH close to neutrality (5.5-6.5) and reduce grain
adhesion (Smith and Love, 1995). The supplemented grains were filled in half or one liter
sterilizable bottles (225 or 450 g/ bottle). Bottles were plugged with non-absorbent cotton and
sterilized in an autoclave at 121ºC for half hour. After cooling, the bottles were inoculated
with pure stock culture by taking a piece of agar with mycelium. The culture and grains were
thoroughly mixed to uniformly distribute the mycelium and were incubated at 25ºC. After all
the grains were fully covered with mycelium, the bottles were used as mother spawn.
3.2.2.2. Inoculation of spawn substrate
Inoculation of spawn substrate in flasks was done by transferring a small amount of agar with
mycelium from stock culture to substrate flask under aseptic condition over a flame. The
mycelium was thoroughly mixed with grains and the flask was then incubated at 25oC in an
incubator. After 7 days of incubation, the content of flasks was shaken to mix the mycelium
uniformly over the grains. The flask was then incubated until all grains become fully covered
with mycelium in about 15 days. Contamination free flasks were used for further
multiplication of spawn (Sara, 2007).
3.2.2.3. Multiplication of spawn from mother culture
Based on the amount of spawn required for the experiment, the spawn substrate was prepared
and sterilized under controlled condition. The substrate flask were inoculated under aseptic
condition in laminar flow cabinet by transferring a small quantity of mycelium covered grains
from master spawn flask to freshly sterilized spawn substrate flasks. Inoculated substrate was
incubated and mycelium was allowed to grow. One flask of master spawn culture was used to
make 30-40 flasks of spawn (Kidane, 2006).
Figure 2 Spawn preparation of oyster mushrooms.
22
3.3. Substrate Preparation and Spawning
Lantana camara, Prosopis juliflora and Parthenium hysterophorus plants or plant parts were
collected from local fields before flowering stage as organic substrates for growing the oyster
mushrooms. The branches of weeds with leaf parts were chopped to 2–4 cm size pieces and
filled in gunny bags and then soaked in 2% aqueous formalin solution for about 18 hours. The
bags, after treatment with formalin solution, were taken out and allowed to drain the excess
solution for 4–6 hours. The soaked pieces of plant materials were then checked for excess
water by pressing between the palms. If water is not dripping, the chopped plant material was
considered as ready for spawning (Sara, 2007). The wet plant materials were then spread on a
clean alcohol swabbed polyethylene sheet and inoculated with spawn prepared at the rate of 3%
on wet basis. The spawn was thoroughly mixed with the substrate and then filled in to plastic
bags (2 kg per bag). Following spawning, the bags were kept about 15 cm apart in a crop
room where the temperature of the room was kept around 25–30oC and humidity 70–90%.
The humidity and temperature range were kept by a heater and by spraying water to the walls
and floors of the cropping room and additionally an instrument known as Data loger was
used. During the cropping period the bags were sprinkled with water twice a day
Figure 3. Wheat (left) and Lantana camara (right) substrate preparation
Figure 4. Parthenium hysterophorus (left) and Prosopis juliflora (right) substrate preparation.
23
3.4. Experimental Design
The experiment was laid out in Completely Randomized Design (CRD) with 4 x 3 factorial
combination of four substrates (Lantana camara, Prosopis juliflora, Parthenium
hysterophorus weeds and wheat straw as control ) and three edible oyster mushroom species
(Pleurotus ostreatus, Pleurotus sajor–caju and Pleurotus florida ) with three replications.
After 25-30 days cultivation, fully matured mushroom species from each substrate were
collected; total yield (in three flush), but the rest like biological efficiency and production rate
was determined on the first flush; mushroom samples were subjected to proximate analysis
(moisture content, crude protein, crude fiber, and ash content).
The treatment combinations were:
No.
Treatment
1
S1M1
2
S2M1
3
S3M1
4
S4M1
5
S1M2
6
S2M2
7
S3M2
8
S4M2
9
S1M3
10
S2M3
11
S3M3
12
S4M3
Where, S1= wheat straw (control)
S2= Prosopis juliflora
M1= Pleurotus ostreatus
S3= Parthenium hysterophorus
M2= Pleurotus sajor–caju
S4= Lantana camara
M3= Pleurotus Florida
24
3.5. Data Collection
3.5.1. Phenological observation
Data on days to the completion of invasion of mycelium on different substrates, appearance of
pin heads formation and days to maturation of fruiting bodies from the day of spawning in
different substrates were recorded.
3.5.2. Yield related parameters
3.5.2.1. Total yield
Weight of fresh fruiting bodies harvested from each substrate block was measured using
sensitive balance. Data on weight of mushroom from each substrate block at first, second and
third flush harvesting stages were recorded separately and their total weight was considered as
total yield which were also used for calculating biological efficiency (Dawit, 1998).
3.5.2.2. Biological efficiency
The weights of each dry substrate and total fresh mushroom weight per bag were recorded
separately and then biological efficiency (BE) of oyster mushrooms in each substrate was
calculated following the formula used by Royse et al., (2004).
BE =
𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑓𝑟𝑒𝑠ℎ 𝑚𝑢𝑠ℎ𝑟𝑜𝑜𝑚 ℎ𝑎𝑟𝑣𝑒𝑠𝑡𝑒𝑑 𝑝𝑒𝑟 𝑏𝑎𝑔
× 100
𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑑𝑟𝑦 𝑠𝑢𝑏𝑠𝑡𝑟𝑎𝑡𝑒 𝑝𝑒𝑟 𝑏𝑎𝑔
3.5.2.3. Production rate
On the basis of biological efficiency on each substrate and the time taken from the spawning
to harvesting, the production rate (PR) of oyster mushrooms in each substrate was calculated
following the method in Dawit (1998).
25
PR = BE/ Time
Where, time is the period in days from the day of spawning to harvesting.
3.5.3. Proximate analysis
Proximate analysis (moisture content, crude protein, crude fiber, and ash content), of the three
edible oyster mushrooms (Pleurotus ostreatus, Pleurotus sajor-caju and Pleurotus florida)
was performed following the methods described by AOAC (1995).
3.5.3.1. Moisture content (%)
The moisture content (MC) of the harvested mushrooms was determined as % moisture loss
in a two stage drying (air dried and oven dried) 2.0 g samples of fresh mushrooms at 70oC to a
stable mass. Then, the dried mass was oven dried (Model: 101–1A, Tianjin Taisite Instrument
Co., Ltd.) at 130oC for 1 hour to a constant weight and the loss in mass was calculated using
the following formula (AOAC, 1995):
𝑀𝑜𝑖𝑠𝑡𝑢𝑟𝑒 𝑐𝑜𝑛𝑡𝑒𝑛𝑡 (%) =
𝑊𝑒𝑡 𝑤𝑒𝑖𝑔ℎ𝑡 − 𝑂𝑣𝑒𝑛 𝑑𝑟𝑦 𝑤𝑒𝑖𝑔ℎ𝑡
× 100
𝑊𝑒𝑡 𝑤𝑒𝑖𝑔ℎ𝑡
Based on moisture percentage, the dry matter percentage was calculated by subtracting each
value from 100 (Singh, 2003).
3.5.3.2. Crude protein (%)
Dried and ground (on a mortar and pestle) samples of oyster mushrooms were analyzed for
crude protein (CP) of each treatment following the methods described by AOAC (1995).
Sample weight (1.0 g) was added into a Kjeldahl digestion flask. Catalyst mixture (Na2SO4
mixed with anhydrous CuSO4 in the ratio of 10:1) of 1.0 g was also added. After addition of
15 mL of H2SO4, digestion flask was placed in the digester and the temperature was brought
26
to 350oC. The flask was removed from the digester (Model: F30100184, SN: 111051, VELP
Scientifica, Europe) and allowed to cool after 5 hours digestion. Then, 30 mL of distilled
water followed by 25 mL of 40% NaOH were added into the digestion flask. The contents
were distilled (Model: F30200191, SN: 111526), Europe) immediately by inserting the
digestion tube line into the receiver flask that contains 25 mL of 4% boric acid solution and
about 150 mL of distillate was collected. Finally, the distillate was titrated by a standard acid
(ca 0.1N HCl). Percentage nitrogen was calculated using the following formula.
𝑁𝑖𝑡𝑟𝑜𝘨𝑒𝑛 (%) =
𝑉𝐻𝐶𝐿 𝑖𝑛 𝑙𝑖𝑡 × 𝑁𝐻𝐶𝐿 (𝑐𝑎. 0.1) × 14.0
× 100
𝑆𝑎𝑚𝑝𝑙𝑒 𝑤𝑒𝑖𝘨ℎ𝑡 𝑖𝑛 𝘨 𝑜𝑛 𝑑𝑟𝑦 𝑚𝑎𝑡𝑡𝑒𝑟 𝑏𝑎𝑠𝑖𝑠 (𝑑𝑏)
Crude protein was determined as follows.
% CP = 6.25 x % N
Urea was used as a control in the analysis.
3.5.3.3. Crude fiber (%)
Crude fiber was determined through digestion of 3.0 g of dried, ground (on a mortar and
pestle) and fat free sample by boiling it in a weak solution of H2SO4 (1.25%) for 30 minutes
followed by boiling it in a weak solution of NaOH (1.25%) for 30 minutes with the addition
of 2–3 pieces of anti–bumping chips. Then, the residue was washed with 25–30 mL of near
boiling water and filtered on to ashless filter paper (90 mm diameter, SN: 1444090, England)
after each wash and dried. The dried residue was transferred to ashing dish and ignited
(AOAC, 1990). The following formula was used to calculate the crude fiber content:
CF (%) =
(𝑊𝑒𝑖𝘨ℎ𝑡 𝑜𝑓 𝑠𝑖𝑙𝑖𝑐𝑎 𝑐𝑟𝑢𝑐𝑖𝑏𝑙𝑒 𝑤𝑖𝑡ℎ − (𝑊𝑒𝑖𝘨ℎ𝑡 𝑜𝑓 𝑠𝑖𝑙𝑖𝑐𝑎 𝑐𝑟𝑢𝑐𝑖𝑏𝑙𝑒
𝐷𝑟𝑦 𝑟𝑒𝑠𝑖𝑑𝑢𝑒 𝑏𝑒𝑓𝑜𝑟𝑒 𝑎𝑠ℎ𝑖𝑛𝘨) 𝑤𝑖𝑡ℎ 𝑎𝑠ℎ 𝑎𝑓𝑡𝑒𝑟 𝑖𝘨𝑛𝑖𝑡𝑖𝑜𝑛)
× 100
𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑓𝑎𝑡 𝑓𝑟𝑒𝑒 𝑠𝑎𝑚𝑝𝑙𝑒 𝑢𝑠𝑒𝑑 (𝘨)(𝑑𝑏)
27
3.5.3.4. Total ash and percent organic matter
Total ash (TA) content was estimated by heating 2g of ground mushroom sample at 550 oC
for 4hrs in a muffle furnace (Model: MF 120, SN: 04–1524, Ankara–Turkey). At this
temperature all the organic matter burns off as CO2 and the remaining matter is inorganic ash
(Singh, 2003). The ash content was calculated according to the following formula:
Total ash (%) = (W3-W1/W2-W1) x 100
Where, W1 = Weight of crucible
W2 = Weight of crucible + Sample
W3 = Weight of crucible + Ash
Based on percentage total ash, percent organic matter was calculated by subtracting each
value from 100.
3.6. Data Analysis
The data collected during the study was subjected to analysis of variance (ANOVA) for
factorial combination of CRD following the procedure described by Gomez and Gomez
(1984) and the least significance difference (LSD) at 5% level of significant was used for the
treatment mean separation. Correlation between the treatments was also computed using SAS
computer program.
28
4. RESULTS AND DISCUSSION
4.1. Effect of Organic Substrates and Mushroom Species on Growth and Yield
4.1.1. Mycelium invasion
Among the different organic substrates, the fastest mycelium invasion of 16.50 days was
recorded for wheat straw (Table 1).The second best substrate in fastest mycelium invasion
was Lantana camara and Parthenium hysterophorus (19.83 days). This might be the nature of
the two invasive weed species with their nutritional composition and chemical composition.
Dried Prosopis juliflora leaves were found to be invaded by inoculated mushroom spawn at
the slowest time (23.83 days). This might be the phenolic compound within the plant and its
antifungal character. Bhatti et al., (1987) reported that the appreciable days to complete
mycelium running of oyster mushroom in different substrates might be due to variations in
their chemical composition and C: N ratio and the data of present study are in agreement with
these findings.
Pleurotus ostreatus, Pleurotus florida and Pleurotus sajor-caju had no significant effect on
mycelium invasion treatments. This could be due to the nature of the Pleurotus species
lignocellulosic enzymes during the invasion time. Prosopis juliflora and wheat straw recorded
significant differences with regard to days to mycelium invasion except Lantana camara and
Parthenium hysterophorus (Table 1).
Interaction of substrates with species of the mushroom was non-significant for days to
mycelium invasion.
29
Table 1: Effect of substrate types on mycelium invasion of Pleurotus species
Substrate
Duration from spawning to mycelium invasion (days)
PO
PF
PS
Lantana camara
19.83c
20.83bc
22.17ab
Parthenium hysterophorus
19.83c
20.37bc
21.17bc
Prosopis juliflora
23.17a
23.83a
23.50a
Wheat straw (control)
16.50d
16.83d
17.33d
CV (%) = 5.67
LSD (5%)= 1.96
PO= Pleurotus ostreatus, PF= Pleurotus florida and PS= Pleurotus sajor-caju
Mean values with in a row followed by the same letter (s) are not significantly different at the
5% probability level.
4.1.2. Pin-head formation
The earliest number of days to pin-head formation after spawning was 20.37days on wheat
straw. The second best earlier pin-head formation was Lantana camara and Parthenium
hysterophorus (19.83 days). Statistically the slowest pin-heading was observed with dried
Prosopis juliflora (23.83 days). This might be the chemical nature (phenolic contents) of the
substrate and also plant type like being herbs and shrubs in which they have different lignin,
cellulose, hemicellulose contents. Cherney, (1989) also reported the inhibitory effect of
phenolic compound on mycelium growth. Significant difference was not observed between
Lantana camara and Parthenium hysterophorus in terms of pin heading. Wheat straw,
Parthenium hysterophorus and Lantana camara were recorded to have led early pin head
formation by Pleurotus species compared to Prosopis juliflora leaves (Table 2).
Pleurotus ostreatus reached pin-head formation earlier than Pleurotus florida and Pleurotus
sajor-caju. This could be due to nature of the fungus (Pleurotus ostreatus) which is also
known as wood mushroom that it may be more competent in degrading the woody substrates
as compared to the other species. The difference observed between the species with regard to
the time taken from spawning to pin head formation was statistically significant (Table 2).
30
Interaction of substrates with species of the mushroom was non-significant for the days Pin
head formation.
Table 2: Effect of substrate types on time taken to pin head formation in Pleurotus species
Substrate
Duration from mycelium invasion to pin head formation
(days)
PO
PF
PS
Lantana camara
24.83cde
24.83cde
26.0bc
Parthenium hysterophorus
23.83e
24.70de
25.17cd
Prosopis juliflora
26.0bc
27.0b
28.83a
Wheat straw (control)
20.37g
21.17fg
22.37f
CV (%) = 2.94
LSD (5%)= 1.22
PO= Pleurotus ostreatus, PF= Pleurotus florida and PS= Pleurotus sajor-caju
Mean values with in a row followed by the same letter (s) are not significantly different at the
5% probability level.
4.1.3. Fruiting body formation
The fastest fruiting body formation after spawning was observed on wheat straw (24.83 days)
followed by Parthenium hysterophorus (26.87 days) this might be the nature of the plant. The
slowest fruiting body formation was recorded on Prosopis juliflora (32.37 days) this could be
the high lignin contents, low cellulose content, low hemicellulose content high phenolic
compounds, high antifungal characteristics and also the existence of the plant to be strong
shrubs although significant difference was not observed between Prosopis juliflora and dried
Lantana camara in terms of fruiting body formation that could be due to the chemical nature
of the two substrates (Table3). The results of this study are in harmony with Robert (1996)
who reported that long period of fruiting body formation is related to high nitrogen content of
a different lignocellulosic substrate (enzymatic activity).
Pleurotus ostreatus had significantly faster (p<0.01) fruiting body formation than Pleurotus
florida and Pleurotus sajor-caju (Table 3).
31
However the interaction between substrates and mushroom species was non-significant in
terms of taken to fruit body formation
Table 3: Effect of substrate types on fruit body formation in three Pleurotus species
Substrate
Duration from pin head formation to fruit body formation
(days)
PO
PF
PS
Lantana camara
29.33b
29.67b
30.53ab
Parthenium hysterophorus
26.87c
29.03b
29.37b
Prosopis juliflora
29.70b
31.0ab
32.37a
Wheat straw (control)
24.83c
25.33c
26.37c
CV (%) = 4.31
LSD (5%)= 2.09
PO= Pleurotus ostreatus, PF= Pleurotus florida and PS= Pleurotus sajor-caju
Mean values with in a row followed by the same letter (s) are not significantly different at the
5% probability level
4.1.4. Yield of oyster mushroom
Harvesting of the mushrooms growing on different substrates was completed in three
consecutive harvesting operations till all the mushrooms were harvested from each substrate.
The yield of Pleurotus ostreatus, Pleurotus florida and Pleurotus sajur-caju in each flush on
each substrate are presented in Fig.5, Fig. 6 and Fig.7, respectively. For Pleurotus ostreatus,
the highest yield was obtained during the first flush from Parthenium hysterophorus, which
therefore, resulted in the highest yield compared to all other substrates. The lowest yield
during the first and third flush was obtained from Prosopis juliflora in case of Pleurotus
sajor-caju, on Fig.7. Whereas the lowest yield in second flush was obtained for Pleurotus
sajor-caju grown on the same plant on Fig.7. In all cases yield showed a declining trend from
the first flush to third flush. This could be due to the decreasing nutrient content of the
substrate consumed by mushroom during growth from one flush stage to the next.
32
900
Mushroom yield in g/2kg
800
700
600
500
1st flush
400
2nd flush
300
3rd flush
200
100
0
WS
LC
PH
PJ
Substrate
Figure 5.Yield of the oyster mushroom (Pleurotus ostreatus) at different flushes on four
substrates (WS= wheat straw, LC= Lantana camara, PH= Parthenium hysterophorus, PJ=
Prosopis juliflora)
600
Mushroom yild in g/2kg
500
400
1st flush
300
2nd flush
3rd flush
200
100
0
WS
LC
PH
PJ
Substrate
Figure 6.Yield of the oyster mushroom (Pleurotus florida) at different flushes on four
substrates (WS= wheat straw, LC= Lantana camara, PH= Parthenium hysterophorus, PJ=
Prosopis juliflora; the unit of first, second and third flushes are in g/2kg dry substrate.)
33
600
Mushroom yild in g/2kg
500
400
1st flush
300
2nd flush
3rd flush
200
100
0
WS
LC
PH
PJ
Substrate
Figure 7. Yield of Oyster mushrooms (Pleurotus sajor-caju) at different flushes (WS= wheat
straw, LC= lantana camara, PH= parthenium hysterophorus, PJ= Prosopis juliflora; the unit
of first, second and third flushes are in g/2kg dry substrate.)
4.1.5. Total yield, biological efficiency and production rate
There was significant (p<0.01) interaction effect (Table 4) between the substrates and
Pleurotus species in terms of the total yield of oyster mushroom. Pleurotus ostreatus on
Parthenium hysterophorus gave significantly higher total yield (1677.45 gm/2kg of dry
substrate) over the other substrates while the lowest yield was obtained from Prosopis
juliflora for Pleurotus sajor-caju (555.43 gm/2kg of dry substrate). This result was to some
extent greater than that of Vats et al. (1994) who grew oyster mushroom on Lantana camara
and wheat straw and reported a yield of 36 kg and 54.8 kg/100kg, respectively, on substrate
dry weight basis.
The highest yield of mushroom grown on Parthenium hysterophorus may be due to the high
nitrogen content, low lignin content(herbs) and narrow C:N ratio (Appendix Table 13) of the
dried Parthenium hysterophorus which provided enough nutrients for the growing
mushrooms but in the case of Prosopis juliflora it was known to be a leguminous plant but the
34
yield it gave was very low perhaps due to the nature of the plant (its high phenolic content),
low cellulose content, high lignin content(because it is shrubs) .
Vijayakhader (2002) reported the inhibitory effect of phenolic compounds leading to lower
fruit body formation and as a result low value had formed. Significant difference was not
observed between Pleurotus ostreatus grown on Lantana camara and Pleurotus sajor-caju
grown on wheat straw.
Table 4: Effect of substrate types on total yield (g /2kg) of Pleurotus species
Substrate
Species
PO
PF
PS
Lantana camara
1222.15d
1023.83g
1098.73f
Parthenium hysterophorus
1677.45a
1178.35de
1106.84ef
Prosopis juliflora
813.51h
753.69h
555.43i
Wheat straw (control)
1538.77b
1325.92c
1234.85d
CV (%) = 3.88
LSD (5%)= 73.77
PO= Pleurotus ostreatus, PF= Pleurotus florida and PS= Pleurotus sajor-caju
Mean values with in a column and row followed by the same letter (s) are not significantly
different at the 5% probability level.
Biological efficiency of various treatment combinations was calculated on the basis of
percentage fresh mushroom production per unit dry weight of substrate. Significant (p<0.01)
interaction effect between the substrates and Pleurotus species on the BE was observed (Table
5). The highest biological efficiency (83.87%) was recorded for Pleurotus ostreatus grown on
dried Parthenium hysterophorus followed by Pleurotus ostreatus grown on wheat straw
(76.94%) this result might be the second highest result in BE because of its high cellulose
content and low lignin content of the substrate (Table5). The BE obtained for Pleurotus
ostreatus grown on dried Parthenium hysterophorus was within the range (68.7-88.4) grows
on paddy straw provided by Kumar (2005). This result was approximate to Alam et al. (2007)
who observed that the BE ranged from 45.21% to 125.70% in case of oyster mushroom on
saw dust and rice straw.
35
Significantly lower biological efficiency (27.77%) was recorded for the Pleurotus sajur-caju
grown on Prosopis juliflora (Table 5). High value of biological efficiency could be observed
for mushrooms grown on narrow C:N ratio substrates such as dried Parthenium
hysterophorus and low value of BE was observed for mushrooms grown on wide C:N ratio,
low lignin and high cellulose substrates relatively. Significant difference in relation to BE was
not observed among Pleurotus ostreatus and Pleurotus florida on Prosopis juliflora plant.
Significant difference was also not observed for Pleurotus sajur caju grown on dried Lantana
camara and Parthenium hysterophorus plant. This might be nature of the plant (Table5).
Table 5: Effect of substrate types on biological efficiency (%) of Pleurotus species
Substrate
Species
PO
PF
PS
Lantana camara
61.11d
51.19g
54.94f
Parthenium hysterophorus
83.87a
58.92de
55.34ef
Prosopis juliflora
40.68h
37.68h
27.77i
Wheat straw (control)
76.94b
66.30c
61.74d
CV (%)=3.88
LSD (5%)=3.69
PO= Pleurotus ostreatus, PF= Pleurotus florida and PS= Pleurotus sajor-caju Mean values
with in a column and row followed by the same letter (s) are not significantly different at the
5% probability level
Significant (p<0.01) interaction effect between different substrates and Pleurotus species was
observed in terms of PR (Table 6). The highest production rates (3.13) were recorded for
Pleurotus ostreatus grown on Parthenium hysterophorus followed by Wheat straw (3.11).
The lowest production rate (0.86) was observed in Pleurotus sajor-caju on Prosopis juliflora.
This could be possibly due to the positive relation in case of PR with nitrogen and the
negative relation with carbon. This observation is in line with Sharma and Madan (1993) who
reported the positive correlation of PR with nitrogen level of the substrates. However,
significant difference was not observed in Pleurotus sajor-caju grown on Lantana camara
and Parthenium hysterophorus in terms of production rate. Pleurotus ostreatus and Pleurotus
florida grown on Prosopis juliflora showed PR values significantly lower than those grown
36
on Parthenium hysterophorus and Lantana camara. Wheat and Lantana camara substrates
were significantly different from one another in terms of their effect on PR regardless of the
mushroom species used (Table 6)
Table 6: Effect of substrate types on production rate of Pleurotus species
Substrate
Species
PO
PF
PS
Lantana camara
2.08d
1.73f
1.80ef
Parthenium hysterophorus
3.13a
2.03de
1.89def
Prosopis juliflora
1.37g
1.22g
0.86h
Wheat straw (control)
3.11a
2.62b
2.35c
CV (%)=7.20
LSD(5%)=0.24
PO= Pleurotus ostreatus, PF= Pleurotus florida and PS= Pleurotus sajor-caju
Mean values with in a column and row followed by the same letter (s) are not significantly
different at the 5% probability level
4.2. Effect of Organic Substrates and Mushroom Species on Quality Parameters
In the present study, moisture content, dry matter percentage, total ash, organic matter, crude
protein, and crude fiber content of oyster mushrooms at harvest were measured to assess
quality of mushrooms grown on different substrates.
4.2.1. Percentage moisture content
Interaction effect of growing substrates and types of species was significant (p<0.01) in
relation to moisture content of the mushrooms (Table 7). The highest moisture content
(92.42%) was recorded in Pleurotus florida grown on wheat straw. This might be the water
holding capacity of the substrate, the nature of mushroom species during the cultivation time.
These results are slightly greater than that of McKellar and Kohrman (1995) and Ortega and
Martinez (1997) who reported a range of 70-90.9% and 89-91% moisture, respectively. The
37
lowest moisture content (85.92) was recorded for Pleurotus sajor-caju on Prosopis juliflora.
This might be the poor nature of the plant in water holding capacity as compared to the other
substrate. The result was to some extent close to the value by Benjamin (2005) who reported
89% for oyster mushrooms grown on paddy straw; similar moisture content (80.0–92.5%)
was reported for Pleurotus sp. grown on different agro wastes (Hamid et al., 1996; Kurtzman,
2005; Ahmed et al., 2009). Moisture content is influenced by mushrooms age, growing
environments, mushroom strains and postharvest environments (Kurtzman, 2005). The
interaction effects between other substrates and Pleurotus species were found variable within
this range.
The high moisture content of the mushrooms observed in this study may suggest that these
mushrooms could be highly perishable as a result of attack by a wide variety of
microorganisms including bacteria. High moisture content has been reported to promote
susceptibility to microbial growth and enzyme activity which accelerates spoilage (Bhupinder
and Ibitwar, 2007).
There was no significant (p<0.01) difference on the Pleurotus species was observed during
the study time
Table 7: The moisture content (%) of Pleurotus species grown on different substrate
Substrate
Species
PO
PF
PS
Lantana camara
88.98cd
86.61e
91.40ab
Parthenium hysterophorus
89.0cd
90.24bc
88.06d
Prosopis juliflora
88.06d
87.96d
85.92e
Wheat straw (control)
89.38c
92.42a
91.76a
CV (%)=0.86
LSD(5%)=1.29
PO= Pleurotus ostreatus, PF= Pleurotus florida and PS= Pleurotus sajor-caju Mean values
with in a column and row followed by the same letter (s) are not significantly different at the
5% probability level
38
4.2.2. Percentage of dry matter
The highest percentage dry matter (14.08%) was recorded from Pleurotus sajor-caju grown
on Prosopis juliflora (Table 8). This may be due to the lowest moisture content of fresh
mushroom (85.92%) grown on Prosopis juliflora (Table 7) The second highest (13.39%)
percentage dry matter of Pleurotus florida on Lantana camara significantly (p<0.01) are
different from other combinations. The lowest significantly (p<0.01) different percentage dry
matter (7.58 %) was observed in Pleurotus florida grown on wheat straw. This may be due to
highest moisture content of mushroom (92.42%) grown on wheat straw and respectively
(Table 7). The dry matter percentage of Pleurotus species found in this study ranged from
7.58 -14.08 % .These results in shared with Bernas et al. (2006) who reported a range of 5.314.8% dry matter percentage. Significant difference was not observed between the Pleurotus
species during the study time. The three species of the mushroom on lantana camara,
Parthenium hysterophorus and Prosopis juliflora also did not differ significantly in terms of
moisture content.
Table 8: The percent dry matter content (%) of Pleurotus species grown on different substrate
Substrate
Species
PO
PF
PS
Lantana camara
11.02bc
13.39a
8.60de
Parthenium hysterophorus
11.0bc
9.76cd
11.94b
Prosopis juliflora
11.94b
12.04b
14.08a
Wheat straw (control)
10.62c
7.58e
8.24e
CV (%)=7.04
LSD(5%)=1.23
PO= Pleurotus ostreatus, PF= Pleurotus florida and PS= Pleurotus sajor-caju Mean values
with in a column and row followed by the same letter (s) are not significantly different at the
5% probability level
39
4.2.3. Total ash content
The interaction effects of growing substrates and type of species were significant (p<0.01) in
relation to total ash content mushrooms (Table 9). The highest total ash content (13.90%) was
recorded for Pleurotus florida grown on Lantana camara while the lowest (6.92%) was for
Pleurotus sajor-caju and Pleurotus ostreatus grown on the Prosopis juliflora substrate. This
value was close to the value mentioned by Ingram (2002) who reported 11.5% ash for
mushrooms grown on wheat straw. Kidane (2006) reported higher ash content (12.32%) for
Pleurotus sajor-caju grown on chat leaves. A number of factors usually influence the
nutritional composition of mushrooms. These factors include growing site, type of substrates,
mushroom type, developmental stages and part of the fungal samples analyzed (Anthony,
2007). This difference could be due to straw type and species type. On the other hand the
lowest ash content result of this study are somewhat close to the Oei (2003) and Dawit (1998)
who reported ash content of 8.80% for oyster mushroom and 7.20% for Pleurotus sajor-caju
respectively. Significant difference was observed among substrates. The differences in ash
contents of the three edible oyster mushrooms used in the present study could be due to the
water used during the cultivation and mushroom species grown (Table 9).
Table 9: The total ash content (%) of Pleurotus species grown on different substrate
Substrate
Species
PO
PF
PS
Lantana camara
11.19b
13.90a
11.26b
Parthenium hysterophorus
11.08b
13.02a
7.69d
Prosopis juliflora
6.92d
9.03c
6.92d
Wheat straw (control)
7.69d
11.06b
11.19b
CV (%)=6.23
LSD(5%)=1.06
PO= Pleurotus ostreatus, PF= Pleurotus florida and PS= Pleurotus sajor-caju Mean values
with in a column and row followed by the same letter (s) are not significantly different at the
5% probability level
40
4.2.4. Percentage organic matter
Organic matter content of oyster mushrooms was significantly (p<0.01) affected by different
substrates and their interaction with Pleurotus species. The highest organic matter (93.08%)
for Pleurotus sajor-caju grown on Prosopis juliflora substrate was recorded and the second
highest organic matter (93.07%) of Pleurotus ostreatus on the Prosopis juliflora was
recorded. whereas the lowest organic matter (86.10%) was recorded for Pleurotus florida
grown on dried lantana plant. The results are somewhat greater than that of Chahal and
Hachey (1990) explained that spent mushroom products grown on maize stalk typically
contain between 40 and 60% OM on a dry weight basis Significant difference was observed
among all the substrates in relation to organic matter (Table 10).This may be due to highest
and the lowest ash content of mushroom grown on dried Lantana camara plant and Prosopis
juliflora respectively (Table 9). Additionally it may be due to the difference in composition of
carbon containing compounds in two substrates and compared to dried lantana leaves, the
carbon in Prosopis may be easily available to growing mycelium and mushrooms.
Table 10: The organic matter content (%) of Pleurotus species grown on different substrate
Substrate
Species
PO
PF
PS
Lantana camara
88.81c
86.10d
88.74c
Parthenium hysterophorus
88.92c
86.98d
92.31a
Prosopis juliflora
93.07a
90.97b
93.08a
Wheat straw (control)
92.31a
88.94c
88.81c
CV (%)=0.70
LSD(5%)=1.06
PO= Pleurotus ostreatus, PF= Pleurotus florida and PS= Pleurotus sajor-caju Mean values
with in a column and row followed by the same letter (s) are not significantly different at the
5% probability level
41
4.2.5. Crude protein
Interaction effect of growing substrates and types of Pleurotus species (Table 11) was
significant (p< 0.01) on crude protein content of edible oyster mushrooms. Crude protein
content (41.48%) of Pleurotus florida grown on Lantana camara was the highest and the
second highest crude protein content (40.51%) of the same species grown on Parthenium
hysterophorus. The cause of high protein contents of mushrooms might be a number of
factors, namely the type of mushrooms, the stage of development, the part sampled, level of
nitrogen available and the location and also the nature of the plant by itself. This values was
slightly similar to the range of (Change and Mshigeni, 2001; Kurtzman, 2005) in which the
Protein on dry matter basis (db) in oyster mushrooms can range 20–40%. The lowest crude
protein (30.11%) was for Pleurotus ostreatus grown on wheat straw that was not significantly
much different from other combinations (Table 11). The lowest crude protein values fall in
the range of values mentioned by Chang et al. (2003) of 26.9–37.2%, 26.6–35.5% and 26.6–
35.6% crude protein content of Pleurotus species grown on various kinds of substrates.
The differences in crude protein could be the high nitrogen content of Lantana camara plant
which contributed towards the higher crude protein content to growing mycelium while wheat
straw supplied less nitrogen and it could be attributed to the efficiency nitrogen utilization by
the species. The positive correlation of high protein content with high nitrogen content of
Lantana substrate implies that nitrogen is essential for synthesis of protein in mushroom
fruiting bodies. This result was in line with Sangwan and Saini (1995) and Ragunathan and
Swaminathan (2003), the protein contents of mushrooms are dependent on biological,
chemical differences and the C:N ratio of substrates. Anthony (2007) also reported that the
protein content of edible mushrooms besides being species/strain specific could also vary with
the growing substrate.
42
Table 11: The crude protein content (%) of Pleurotus species grown on different substrate
Substrate
Species
PO
PF
PS
Lantana camara
32.33c
41.48a
37.63ab
Parthenium hysterophorus
32.76c
40.51a
31.60c
Prosopis juliflora
30.87c
32.60c
31.77c
Wheat straw (control)
30.11c
32.69c
33.84bc
CV (%) =7.06
LSD (5%)= 4.04
PO= Pleurotus ostreatus, PF= Pleurotus florida and PS= Pleurotus sajor-caju Mean values
with in a column and row followed by the same letter (s) are not significantly different at the
5% probability level
4.2.6. Percentage crude fiber
The crude fiber content of edible oyster mushrooms was significant (p<0.01) due to the
interaction effects of growing substrates and type of mushroom species (Table 12). Crude
fiber content (12.73%) of Pleurotus sajor-caju grown on wheat straw was the highest whereas
the second highest crude fiber content (8.87%) of Pleurotus florida grown on wheat straw.
These values are close to the value mentioned by Crisan and Sands (1978) who reported
13.3% crude fiber for cultivated edible mushrooms and Kidane (2006) who reported 10.6%
crude fiber content for Pleurotus sajor–caju. The lowest Crude fiber (5.19%) of Pleurotus
ostreatus on Prosopis juliflora was formed. This result was close to the range of the Obodai
(1992) reported fiber values of 6.5–16.3% for Pleurotus species. Over all the crude fiber
content in different kinds of oyster mushroom might be dependent on the harvesting time of
the particular mushroom species. Significant differences in crude fiber content of mushroom
were observed within the different substrate in this study. The difference in crude fiber
content could be attributed to the variation in the nutritional compositions of substrates and
the type of Pleurotus species grown.
43
Table12: The percent crude fiber content (%) of Pleurotus species grown on different
substrate
Substrate
Species
PO
PF
PS
Lantana camara
6.67cd
5.75d
7.54bc
Parthenium hysterophorus
8.56b
6.52cd
7.93bc
Prosopis juliflora
5.19d
5.65d
7.71bc
Wheat straw (control)
8.61b
8.87b
12.73a
CV (%)=12.16
LSD(5%)=1.57
PO= Pleurotus ostreatus, PF= Pleurotus florida and PS= Pleurotus sajor-caju Mean values
with in a column and row followed by the same letter (s) are not significantly different at the
5% probability level
4.3. Correlation Among Parameters
4.3.1. Correlation between mushroom yield parameters
Results of the simple linear correlation analysis (Table 13) showed that TY had a high
positive and significant correlation with BE (r = 1.0***) and PR (r = 0.98***). This indicated
that high TY was essential for biological efficiency and production rate to increase but low
yield had negative effect on the yield parameters. A positive and significant correlation
between the days of mycelium invasion such as on set of pin head formation (r =0.87***),
onset of fruiting body formation (r = 0.79***) and also pin head formation (0.92***) had a
positive correlation with day of fruit formation which indicated a close relationship between
mycelium invasion and other yield parameters. The TY had strong negative correlation with
Phenological parameters such as days of mycelium invasion (r = -0.73***), days of pin head
formation (r = -0.79***) and also days of fruit body formation (r = -0.77***) in other words
BE and PR was also negatively correlated. A result from main effect of this study indicates
that Phenological parameters have no direct influence on the other yield parameters.
44
4.3.2. Correlation between mushroom quality parameters
Results of the simple linear correlation analysis (Table 14) showed a negative and significant
correlation of MOC with DM (r = -1.0***) and non significant with TA (r = 0.31ns), OM (r =
-0.31ns) and CP (r = 0.11ns) but on the case of CF (r = 0.49**) it was significant. DM with
TA (r = - 0.31ns), OM (r = 0.31ns) and CP (r = -0.11ns) was significant but in the case of CF
(r = -0.49**) it was negatively correlated. A negative and significant correlation of TA with
OM (r = -1.0***) was formed but in case of CP (r = 0.69***) it was positive and significant
and in CF (r =0.04ns) it was non-significant. In the case of OM it was negatively correlated
with CP (r = -0.69***) and CF (r = -0.04ns). This was may be because one has no influence
on the other. But crude protein with crude fiber (r =-0.18ns) was non-significant.
Table 13.Correlation coefficient for different yield parameters of mushroom
DMI
DPF
DFF
BE
PR
DMI
1
DPF
0.87***
1
DFF
0.79***
0.92***
1
BE
-0.73***
-0.79***
-0.77***
1
PR
-0.78***
-0.86***
-0.87 ***
0.98***
1
TY
-0.73***
-0.79***
-0.77***
1.0***
0.98***
TY
1
*** indicate significant difference at 0.001 probability levels, respectively. DMI = Days to
mycelium invasion, DPF= Days to pin-head formation FBF= Days to fruiting body formation,
BE=biological efficiency, PR=production rate and TY= Total yield
45
Table 14.Correlation coefficient for different quality parameters of mushrooms
MOS
DM
TA
OM
CP
MOS
1
DM
-1.0***
1
TA
0.31ns
-0.31ns
1
OM
-0.31ns
0.31ns
-1.0***
1
CP
0.11ns
-0.11ns
0.69***
-0.69***
1
CF
0.49 **
-0.49**
0.04ns
-0.04ns
-0.18ns
CF
1
***, ** and ns indicate significant difference at 0.001, 0.05 and non significant probability
levels, respectively. % MOS= percentage moisture content, %DM= percentage dry matter,
%TA= Percentage total ash, %OM= Percentage organic matter, % CP = Percentage crude
protein and %CF = Percentage crude fiber.
46
5. SUMMARY AND CONCULUSION
5.1. Summary
Growth, yield and quality of mushrooms are substantially influenced by the substrate type on
which they are grown. In view of this, the present research work was conducted at the
Mushroom Research, Production and Training Center of Haramaya University during the year
2011-2012. A factorial experiment involving three oyster mushroom species (Pleurotus
ostreatus, Pleurotus florida and Pleurotus sajor-caju) and four substrates (wheat straw,
parthenium hysterophorus, lantana camara and Prosopis juliflora) was undertaken in a
complete randomized design with three replications.
The fastest mycelium formation was recorded for wheat straw while slowest mycelium
formation was on dried Prosopis juliflora. The earliest pin head were formed on wheat straw
while dried Prosopis juliflora recorded the slowest pin head formation. The earliest fruit
bodies were formed on wheat straw while dried Prosopis juliflora recorded the slowest
fruiting body. Pleurotus ostreatus treated substrates had significantly (p<0.01) earlier
invasion by mycelium, pin head and fruit bodies formation than Pleurotus florida and
Pleurotus sajur-caju treatments. The interaction effects revealed Pleurotus ostreatus on
Parthenium hysterophorus yet again Pleurotus ostreatus on wheat straw gave significantly
(P< 0.01) higher TY respectively, the lowest yield was obtained from Prosopis juliflora in the
case of Pleurotus sajor-caju. The higher BE was recorded for Pleurotus ostreatus grown on
dried Parthenium hysterophorus and Pleurotus ostreatus treated wheat straw was found to be
the second for the BE compared to all other treatment. Significantly (p<0.01) lower BE was
recorded for the mushrooms grown on Prosopis juliflora. Similarly the higher PR was
recorded for Pleurotus ostreatus grown on Parthenium and Wheat straw. While the lowest PR
was observed in Pleurotus sajor-caju on the dried Prosopis juliflora
Interaction effect of growing substrates and types of species was significant (p<0.01) in
relation to PMC of the mushrooms. The highest PMC obtained was recorded in Pleurotus
florida species grown on wheat straw. The lowest PMC for Pleurotus sajor-caju on the
47
Prosopis juliflora was recorded. PDM of Pleurotus sajor-caju grown on Prosopis juliflora
was the highest. The second highest PDM of Pleurotus florida on Lantana camara
significantly (p<0.01) are different from other combinations. The lowest significantly
(p<0.01) different PDM was observed in Pleurotus florida grown on wheat straw. The highest
TA content was recorded for Pleurotus florida grown on Lantana camara while the lowest
was for Pleurotus sajor-caju grown on the Prosopis substrate. Organic matter for Pleurotus
sajor-caju grown on Prosopis juliflora substrate was highest and the second highest OM of
Pleurotus ostreatus on the Prosopis plant was recorded. Whereas the lowest OM was recorded
for Pleurotus florida grown on dried Lantana plant. CP content of Pleurotus florida grown on
Lantana camara was the highest and the second highest CP content of the same species
grown on Parthenium hysterophorus .The lowest CP was for Pleurotus ostreatus grown on
wheat straw that was not significantly much different from other combinations. Significantly
(p< 0.01) of CF content of Pleurotus sajor-caju grown on wheat straw was the highest the
second highest CF content of Pleurotus florida grown on wheat straw. The lowest of
Pleurotus ostreatus on Prosopis was formed.
Results of the simple linear correlation analysis showed that TY had a high positive and
significant correlation with BE (r = 1.0***) and PR (r = 0.98***). A positive and significant
correlation between the days of mycelium invasion such as on set of pin head formation (r
=0.87***), onset of fruiting body formation (r = 0.79***) and also pin head formation
(0.92***) had a positive correlation with day of fruit formation which indicated a close
relationship between mycelium invasion and other yield parameters. The TY had strong
negative correlation with Phenological parameters such as days of mycelium invasion (r = 0.73***), days of pin head formation (r = -0.79***) and also days of fruit body formation (r =
-0.77***). A result from main effect of this study indicates that Phenological parameters have
no direct influence on the other yield parameters.
Results of the simple linear correlation analysis showed a negative and significant correlation
of MOC with DM (r = -1.0***) but on the case of CF (r = 0.49**) it was positive and
significant. DM with CF (r = -0.49**) it was significant and negatively correlated. A negative
and significant correlation of TA with OM (r = -1.0***) was formed but in case of CP (r =
0.69***) it was positive and significant. In the case of OM it was negatively correlated with
48
CP (r = -0.69***) and CF (r = -0.04ns). This was may be because one has no influence on the
other.
5.2. Conclusions
In general, results of the present study showed that organic residues having wide C:N ratio
reduced the yield and quality of mushrooms. On the other hand, substrate residues with
narrow C:N ratio such as parthenium and lantana had positive correlation with yield and
quality of mushrooms but with some risk of relatively higher contamination apparently due to
the presence of readily available nutrients. The present study was conducted with a limited
number of substrates. However there is a lot of scope for conducting similar studies on
different substrates which are easily available locally to the farmers interested to grow
mushrooms under different agro climatic conditions across Ethiopia.
5.3. Recommendation
a. Invasive weed species with other supplements (wheat bran, Calcium sulfate, Nitrogen
compound) – to improve yield of oyster mushroom.
b. Further studies on the nutritional and chemical contents of invasive weed species for the
improvement of nutritional quality within the oyster mushroom.
49
6. REFERENCE
Abdullah, A.Y., B.Y. Abddel hafes, 2004. Inclusion of Prosopis juliflora pods in finishing
Awassi lamb diets. In: Proc. 11th AAAP Animal Science Congress, vol. 2, pp. 373–375.
Adamovic, M.G., I.S. Grubic, T.R. Milenkovc, R.K. Jovanovic and L. S. Oeivic, 1998. The
bio-degradation of wheat straw by Pleurotus ostreatus mushrooms. Animal Feed Science
Technology l71: 357-362
Aggelis, G., D. Iconomou, M. Christou, D. Bokas, S. Kotzailias, G. Christou, V. Tsagou, S.
Papanikolaou, 2003. Phenolic removal in a model olive oil mill wastewater using Pleurotus
ostreatus in bioreactor cultures and biological evaluation of the process. Water Research 37:
3897-3904.
Ahmad, I., 1986. Some studies on oyster mushrooms (Pleurotus species) on waste materials
of corn industry. MSc Thesis. Department of Plant Pathology, Faisalabad. pp. 25-50
Ahmed, S.A., J.A. Kadam, V.P. Mane, S.S. Patil, & M.M.V. Baig, 2009. Biological
efficiency and nutritional contents of Pleurotus florida (Mont.) singer cultivated on different
agro-wastes. Nature and Science, 7: 44–48.
Alam, N., SMR. Amin. and NC. Sarker, 2007. Efficacy of five different growth regulators on
the yield and yield contributing attributes of Pleurotus ostreatus (Jacquin ex Fr.) Kummer.
Bang. J. Mush. 1 (1): 51-55.
Anthony, M.M., 2007. Proximate and nutrient composition of three types of indigenous edible
wild mushrooms grown in Tanzania and their utilization prospects. African Journal of Food,
Agriculture, Nutrition and Development, 7(6): 2–9.
AOAC (Association of Official Analytical Chemist), 1990. Method 962.09. Official Methods
of Analysis of AOAC International. pp. 80–81. (15th eds.). Arlington, Virginia U.S.A.
AOAC (Association of Official Analytical Chemist), 1995. Official Methods of Analysis, 14th
Edition, Washington, DC.
AUA (Alemaya University of Agriculture), 1996. Proceedings of the 13th annual research and
extension review meeting. Alemaya University of Agriculture, February 26–28, Alemaya,
Ethiopia.
Bano, Z., M.N. Shasirekha and S. Rajarathnam, 1993. Improvement of the bioconversion and
biotransformation efficiencies of the oyster mushroom (Pleurotus sajor-caju) by
supplementation of its rice straw with oil seed cakes. Enzyme and Microbial Technology 15:
985–989.
Baysal, E., H. Peker, M. Kemal, A. Temiz, 2003. Cultivation of oyster mushroom on waster
paper with some added supplementary materials. Bioresour. Technol. 89: 95-97.
50
Bernardi, E., E. Minotto, J.S. Nascimento, 2008. Aproveitamento deresíduo decurtume como
Suplemento no cultivo de Pleurotus ostreatus. Arq. Inst. Biol. 72: 243-246.
Bernas, E.T., G.B. Jawaraska and Z.M. Lisiewska, 2006. Edible mushroom as a source of
valuable nutritive constituents. Acta Sci. Pol. Technol. Aliment 5(1): 5-20.
Benjamin, B.W., 2005. Mushroom Production. Oyster raw http://www.nutrition data. Com
/facts. B0001-01czonc.htm.
Bhatti MA, Mir FA, M. Siddiq, 1987. Effect of different bedding materials on relative yield of
oyster mushroom in the successive flushes. Pak. J. Agri. Resear. 8(3): 256-259.
Bhupinder, K. and B.B. Ibitwar, 2007. Mushroom cultivation and processing. Science Tech
Entrepreneur. Senior Food Technologist, Department of Food Science and Technology,
Punjab Agricultural University, Ludhiana – 141004
Binding, G.J., 1972. Mushrooms, Nature’s Major Protein Food. Thomsons Publishers Ltd.
Northampton Shire, G.B. 64p.
Biradar D.P, K.S. Shivakumar, S.S. Prakash, T. Pujar, 2006. Bionutrient potentiality of
Parthenium hysterophorus and its utility of green manure in rice ecosystem. Karnataka J.
Agric. Sci. 19: 256-263.
Bist, N.S and S.K. Harsh, 1983. Cultivation of Pleurotus ostreatus (Jacq.) Fr. by utilizing
Lantana camara and waste paper. Agric. Wastes, 11: 99-103.
Bist, N.S and S.K. Harsh, 1985. Biodegradation of Lantana camara and waste paper to
cultivate Agaricus bisporus singer. Aric. Wastes, 12: 167-172.
Bobek, P., L. Ozdin and S. Galbavy, 1998. Dose- and time-dependent hypocholesterolemic
effect of oyster mushroom (Pleurotus ostreatus) in rats. Nutrition 14: 282-286.
Bononi, V.L., M. Capelari, R. Mazieiro, SFB. Trufem, 1999. Cultivo de Cogumelos
Comestíveis. Ícone. São Paulo, Brazil. 206 pp.
Brenneman, J.A., M.C. Guttman, 1994. The edibility and cultivation of the oyster mushroom.
Am Biol Teacher; 56: 291-293.
Cangy, C and A. Peerally, 1995. Studies of Pleurotus production on sugarcane bagasse.
African Journal of Mycology and Biotechnology 3: 67–79.
Chahal, D.S. and J.M. Hachey, 1990. Use of hemicellulose, cellulose and degradation of
lignin by Pleurotus sajor-caju grown on corn stalk. A-C-S-Symp-Ser –Am-Chem-Soc. 433:
301- 310
Chang S.T., 1972. Studies of Pleurotus production on sugarcane bagasse. African Journal of
Mycology and Biotechnology 3: 67–79
51
Chang, S.T. and W.A. Hayes, 1998. The Biology and Cultivation of Edible Mushrooms.
Academic press. Inc. London.
Chang, S.T., 1999. World production of cultivated edible and medicinal mushrooms in 1997
with emphasis on Lentinus edodes (Berk.) Sing. In China. International Journal of Medicinal
Mushrooms, 1: 291-300
Chang, R., 1996. Functional properties of Edible Mushrooms. Nutrition Reviews, 54: 591–
593.
Chang, S.T. and P.G. Miles, 1989. Part I—Mushroom science. Edible mushroom and their
cultivation. Boca-Raton, USA: CRC Press.
Chang, S.T. & K.E. Mshigeni, 2001. Mushroom and their Human Health: Growing
Significance as Potent Dietary Supplements. Pp.1– 79. Windhoek: University of Namibia.
Chang, S.T.; O.W. Lav and K.Y. Cho, 2003. The cultivation and nutritional value of
Pleurotus sajor–caju. European Journal of Applied Microbiology and Biotechnology, 15(2):
60-72.
Channappagoudar B.B, N.R. Biradar, J.B. Patil, C.A.A. Gasimani, 2007. Utilization of weed
biomass as an organic source in sorghum. Karnataka J Agric. Sci. 20: 245-248.
Cherney, J.H., K.S. Anliker, K.A. Albrecht and K.V. Wood, 1989. Soluble Phenolic
monomers in forage crops. Journal of Agriculture and food Chemistry 37: 345-350.
Crisan, E.V. and A. Sands, 1978. Nutritional value. In: S.T. Chang and W.A. Hayes (Eds).
The Biology and Cultivation of Edible Mushrooms. London, Academic Press Inc. pp137–165.
Croan, S.C., 2000. Conversion of wood waste value-added products by edible and medicinal
Pleurotus (Fr.) P. Karst Species (agaricales S.I., Basidiomycetes). Int. J. Med. Mush. 2: 773–
780.
Cohen, R., L. Persky and Y. Hadar, 2002. Biotechnological applications and potential of
wood degrading mushrooms of the genus Pleurotus. Applied Microbiology and
Biotechnology. 8: 582–594.
Das, N. and M. Mukherjee, 2007. Cultivation of Pleurotus ostreatus on weed plants. Bio
Resource Technology 98: 2723 – 2726.
Davis, R.A. and B.J. Aegerter, 2006. Edible mushroom cultivation. Department of Plant
Pathology, University of California. http://somamushroom.org/speakers/dais.htm.
Dawit, A., 1998. Mushroom Cultivation; a Practical Approach. Berhanena Selam Printing
Enterprise, Addis Ababa.
52
De Barros, N.A.M.T, C.A.G. Bai and F.C.E Fonseca, 1988. Use of Prosopis juliflora (Sw)
DC and cassava (Manihot utilissima Pohl) for confined sheep feeding during the dry season.
In: Mario A. Habit and Julio C. Saavedra (eds): The current state of knowledge on Prosopis
juliflora. 1988. II International Conference on Prosopis. 25th-29th August 1986, Recife,
Brazil. Food and Agriculture Organization of the United Nations (FAO), Plant Production and
Protection Division.
Duke, A. James, 1983. Prosopis juliflora DC In: Handbook of Energy Crops. Purdue
University Center for New Crops and Plant Products. Version of 1998-JAN-08. Retrieved
2008-MAR-19.
FAO (Food and Agriculture Organization), 1980. The state of Food Insecurity in the World.
2nd ed. Viale della Terme di caracalla, 00100. Rome, Italy.
FAO (Food and Agriculture Organization), 2006. The state of food insecurity in the world.
FAO organization Report. FAO, Rome, Italy.
Felker, Peter and James Moss, 1996. Prosopis: semi arid fuel wood and forage: tree building
consensus for the disenfranchised. A workshop. 13th –15th march, 1996. US National
Academy of Science.
Fianu, F.K., R.K. Assoku, P. Anumel, 1981. Poisonous weeds in pastures: experimental
studies in animals with Tephrosia purpurea (L) Pers. Bull. Anim. Health. Prod. Str. 29: 341–
348.
Fountoulakis, M.S., S.N. Dokianakis, M.E. Kornaros, G.G. Aggelis, G. Lyberatos, 2002.
Removal of phenolics in olive mill wastewaters using the white-rot fungus Pleurotus
ostreatus. Water research 36: 4735- 4744.
Gadd, G.M., 2001. Fungi in bioremediation. Cambridge: Cambridge University Press.
Tropical soil seed banks. In: Leck, M.A., Parker, V.T., Simpson, R.L. (Eds.), Ecology of Soil
Seed Banks. Academic Press Inc., San Diego, pp. 149–363.
Gong, J., K.C. Chan, S.W. Chiu, 2006. Toxicities of DDE on wheat and bioremediation of
DDE by fungus Pleurotus pulmonarius. Hum. Ecol. Risk Assess. 12: 221–235.
Geml, J., P. Labuschagne, D.J. Royse, 2001. Oyster mushroom production on three
continents: An overview of cultivation in Hungary, South Africa and the United States. Mush.
News. 49 (2): 4–13.
Gerasimenya, V.P., O.V. Efremenkova, O.V. Kamzolkina, I.A. Bogush, I.V. Tolstych and
V.A. Zenkova, 2002. Antimicrobial and antitoxical action of edible and medicinal mushroom
Pleurotus ostreatus (Jacq.Fr.) Kumm. Extracts. International Journal of Medicinal
Mushrooms 4: 127- 132.
Gooden, B., K. French, P.J. Turner, 2009. Invasion and management of a woody plant,
Lantana camara L., alters vegetation diversity within wet sclerophyll forest in southeastern
Australia. Forest Ecology and Management 257: 960–967.
53
Gomez and K.A. Gomez, 1984. Statistical Procedures for Agricultural Research. Jone Willy
and Sons. Inc, New York.
Gunde-Cimerman, N., 1999. Medicinal value of the genus Pleurotus (Fr.) P. Darst.
(Agaricales s.l., Basidiomycetes). International Journal of Medicinal Mushrooms 1: 69-80.
Habit, M.A., J.C. Saavedra, 1988. The Current State of Knowledge on Prosopis juliflora.
FAO, Plant Production Division, Rome
Hami, H., 1990. Cultivation of oyster mushroom on sawdust of different woods. An M.Sc.
Thesis, University of Agriculture, Faisalabad, Pakistan.
Hamid, A.K., F.H. Shah & M.A. Qudeer, 1996. Nutritional composition of oyster
mushrooms. Pakistan Journal of Biochemistry, 6: 56–67.
Hassan El-Sayed Embaby and Sayed Mohamed Mokhtar, 2011. Chemical Composition and
Nutritive Value of Lantana and Sweet Pepper Seeds and Nabak Seed Kernels. Journal of
Food Science 76(5) 736-741.
Harris, P.J.C., N.M. Pasiecznik, S.J. Smith, J.M. Billington, L. Ramirez, 2003. Differentiation
of Prosopis juliflora (Sw.) DC. and P. pallida (H. and B. ex. Willd.) H. B. K. using foliar
characters and ploidy. For. Ecol. Manage. 108:153–164.
Hartley, R.D., 1987. The chemistry of lignocellulosic materials from agricultural wastes in
relation to processes for increasing their biodegradability. In Degradation of Lignocellulosic
in Ruminants and in Industrial processes. Eds J.M. Van Der Meer, B.A. Rijkens and M.P.
Ferranti. Elsevier Applied Science, London, pp. 3-12.
Hattaka, A., 1994. Lignin -modifying enzymes from selected white rot fungi: production and
role in lignin degradation. FEMS Microbiol Rev; 13:125–35.
Hopkins, M., 2007. Cooking up a smoky solution, Nature News, doi: 10.1038/ News 0708131, News.
Ingram, S.P., 2002.The real nutritional value of fungi. http://www World of fungi. Org/mostly
medical/ Stephanie ingram /Nutritional value htm as retrieved on Feb 2006.
Jandaik, C.L and Goyal, S.P., 1995. Farm and farming of oyster mushroom (P. species). pp.
72-78. In: R.P.Singh and H.S. Chaube. Mushroom Production Technologies, G B Pant
University of Agriculture and Technology, Pantngear, 263145, U.P. India.
Jose, N., T.A. Ajith and K.K. Jananrdhanan, 2002. Antioxidant, anti- inflammatory, and
antitumor activities of culinary-medicinal mushroom Pleurotus pulmonarius (Fr.) Quel.
(Agaricomycetideae). International Journal of Medicinal Mushrooms 4: 329-335.
Kalberer, P.P., 1974. The cultivation of Pleurotus: Experiments to elucidate the influence of
different culture conditions on the crop yield. Mushroom Science, 9: 653–662.
54
Kang, S.W., 2004. Introduction to oyster mushroom. In: Oyster Mushroom Cultivation.
Mushroom Growers Handbook.1. Mush world.
Kidane, M., 2006. Effect of organic substrate and their pasteurization methods on yield and
quality of oyster mushroom (Pleurotus sajor caju). A M.Sc. Thesis Presented to the School of
Graduate Studies of Haramaya University. 24-25 pp.
Kues, U. and Y. Liu, 2000. Fruiting body production in basidiomycetes .Appl. Microbiol.
Biotechnol. 54: 141-152.
Kumar, G.B., 2005. Mushroom cultivation method. http:// karlmon.fjsyj-net/article- print asp
article I234 .
Kurtzman, R.H. Jr., 2005. A review mushrooms: sources for modern western medicine.
Micologia Aplicada International, 17: 21–33.
Laborde, J., G. Lanzi, B. Francescutti, E. Giordani, 1993. Mushroom Biology and Mushroom
Products. pp. 93–113. In: Chang, S.T., J.A. Buswell, S.W. Chiu (Eds.). Indoor composting:
General principles and large scale development in Italy. The Chines University Press, Hong
Kong.
Lau, K. L., Tsang, Y.Y., Chiu, S.W., 2003.Useofspentmushroomcomposttobiore- mediation
PAH-contaminated samples. Chemosphere 52:1539-1546.
Libnernonnecke, I.B., 1989. Vegetable Production. Van Nostrand Rein hold, New York.
Phytoremediation: biological cleaning of a polluted environment. Rev. Environ. Health 19:
63–82
Mattilda, P., K. Könkö, M. Eurola, J.M. Pihlava, J. Astola, L. Vahteristo, 2001. J Agric Food
Chem 49: 2343–2348.
Martı´nez-Carrera, D., G. Guzma´n, C. Soto, 1985. The effect of fermentation of coffee pulp
in the cultivation of Pleurotus ostreatus in Mexico. Mush. Newsletter Tropics 6: 21–28.
Martı´nez-Carrera, D., 1989. Simple technology to cultivate Pleurotus on coffee pulp. Mush.
J. Tropics 7: 13–23.
Mckeller, R.L and R.E. Kohrman, 1995. Oyster mushroom production. J. Agr. Food Chem.
24: 549-570.
McNeely, J. A., Mooney, A. H., Scheip, E. L. and K. J. Waage, 2001. A Global strategy on
Invasive Alien species, IUCN Gland Switzerland and Cambridge, UK, in collaboration with
GISP.50p.
Miles, P.G., S.T. Chang, 1997. Mushroom Biology, Concise Basics and Current
Developments; World Scientific: Singapore
55
Muzzarelli, R.A.A., 1999. Clinical and biochemical evaluation of chitosans for
hypercholesterolemia and overweight control. In: Chitin and Chitinases. Eds.: P. Jollies,
Narayanmurti (1955). Indian For. Bull; N.S., No. 207, 41.
Nicola, K., 2003. Hands on Farming Fungus. http:// www. tve.org./ho/doc.cfm?aid=996.
Nigam, S.K., V.N. Sharma and K.N. Kaul, 1977. New compounds from Lantana camra. Res
.indus. New Delhi, 2: 194-195.
Obodai, M., 1992. Comparative studies on the utilization of agricultural waste by some
mushrooms (Pleurotus and Volvariella species). M. Phil., the University of Ghana, Legon.
Oei, P., 1996. Mushroom Cultivation: Techniques, Species and Opportunities for Commercial
Applications, Amsterdam. The Netherlands.
Oei, P., 2003. Mushroom Cultivation: with Special Emphasis on Appropriate Techniques for
Developing Countries. Tool Publication, Leiden, the Netherlands.
Olivieri, G., A. Marzocchella, P. Salatino, P. Giardina, G. Cennamo, G. Sannia, 2006. Olive
mill waste water remediation by means of Pleurotus ostreatus. Biochemical Engineering
Journal 31: 180-187.
Ortega, G.M. and E.O. Martinez, 1997. Cultivation of Mushroom. World Journal of
Microbiology and Biotechnology 8: 308-320
Pandey, A., C.R. Soccol, P. Nigam, D. Brand, R. Mohan, S. Roussos, 2000. Biotechnological
potential of coffee pulp and coffee husk bioprocesses. Biochem. Eng. J. 6: 153–162.
Pasieczink, N.M., 2001. The Prosopis juliflora–Prosopis pallida Complex: A Monograph.
HDRA, Coventry, UK.
Pettipher, G.L., 1987. Cultivation of the oyster mushroom (Pleurotus ostreatus). Journal of
the Science of Food and Agriculture, 41: 259–265
Prem Kishor, A.K. Ghosh, Surendra Singh and B. R. Maurya, 2010. Potential use of
parthenium (Parthenium hysterophorus L.) in Agriculture. Asian Journal of Agricultural
Research 4 (4): 220-225
Quimio, T.H., S.T. Chang and D.J. Royse, 1990. Technical Guidelines for Mushroom
Growing in the Tropics. FAO Plant Production and Protection Paper, 106. Rome.
Ragunathan, R. and K. Swaminathan, 2003. Nutritional status of Pleurotus species grown on
various agro–wastes, Food Chemistry, 80(3): 371–375.
Robert, K., 1996. Spawn running and enzymatic activity. Brazilian Archive of Biology and
Technology 55(2): 455-468.
56
Royse, D.J., 2002. In Xuence of spawn rate and commercial delayed release nutrient levels on
Pleurotus cornucopiae (oyster mushroom) yield, size and time to production. Appl. Microbiol.
Biotechnol. 58: 527–531.
Royse, D.J., T.W. Rhodes, S.Ohga, J.E. Sanchez, 2004. Yield, mushroom size and time to
production of Pleurotus cornucopiae (oyster mushroom) grown on switch grass substrate
spawned and supplemented at various rates. Bioresource Technology 91: 85–91.
Sadler, M., 2003. Nutritional properties of edible fungi. Nutrition bulletin, 28(3): 305–308.
Sanchez, A., F. Ysunza, M. Beltran-Gracia and M. Esqueda, 2002. Biodegradation of
viticulture wastes by Pleurotus: a source of microbial and human food and its potential use in
animal feeding. Journal of Agricultural and Food Chemistry 50: 2537–2542.
Sangwan, M.S. and L.C. Saini, 1995. Cultivation of Pleurotus sajor–caju (Fr.). Singer on
Agro industrial Wastes. Mushroom Research, 4: 33–34.
Sara, M., 2007. Effect of spawn substrates and their inoculation rates on the yield and quality
of oyster mushroom (Pleurotus florida). An M.Sc. Thesis Presented to the School of Graduate
Studies of Haramaya University. 22-23 pp.
Seifu W., 1990. Parthenium hysterophorus L., a recently introduced noxious weed to
Ethiopia. A preliminary reconnaissance survey report on Eastern Ethiopia. East Harerge,
Ministry of Agriculture, Ethiopia.
Semple, K.T., Reid, B.J., Fermor, T.R., 2001. Impact of composting strategies on the
treatment of soils contaminated with organic pollutants: a review. Environ- mental Pollution
112: 269-283.
Sharma, S. and M. Madan, 1993. Microbial protein from leguminous and non leguminous
substrates. Acta Biotechnology 13(2): 131-139.
Sharma, O. P., H.P.S. Makkar, R.K. Dawra, 1988. A review of the noxious plant Lantana
camara. Toxicon 26: 975–987.
Sharma, O.P., R.K. Dawra, and V. Pattabhi, 1991. Molecular structure, polymorphism and
Toxicity of lantadene A, the pentacyclic triterpenoid from the hepatotoxic Lantana camara. J.
Biochem. Toxicol., 6: 57-63.
Shah, Z.A., M. Ashraf and Ch. M. Ishtiaq, 2004. Comparative study on cultivation and yield
performance of oyster mushroom (Pleurotus ostreatus) on different substrates (Wheat straw,
Leaves, Saw dust). Pakistan Journal of Nutrition 3(3): 158-160.
Shasho Megerssa, 2004. A brief review on use of mushrooms in Ethiopia. Forestry research
news letter 1 (1): 11-12.
Sheeja, R.Y., T. Murugesan, 2002. Studies on biodegradation of phenol using response
surface methodology. J Chem Technol Biotechnol; 77:1219–1230.
57
Shukla, P.C., P.M .Talpada, and M.B. Pande, 1984. Agro-Industrial byproducts as livestock
feed. Prosopis juliflora pods a new cattle feed source. Technical Bulletin, I.C.A.R. Animal
Nutrition Department, Gujarat Agricultural University, Anand, - 388110, India.
Simpson, B.B., 1977. Mesquite, its biology in two desert scrub ecosystems. Dowden,
Hutchinson & Ross, Inc. Stroudsburg, PA.
Singh, A.K., H.K. Sharma, P. Kumar and B. Singh, 1999. Physicochemical changes in white
button mushroom (Agaricus bisporus) at different drying temperatures. Mushroom Research
8(2): 27–30.
Singh, K.L., 2003. Animal nutrition laboratory manual cum record. Department of Animal
Nutrition University of Agricultural Sciences. Hebbal Bangalore, 560024.
Singh, R.P. and H.S. Chaube.1995. Mushroom Production Technologies: Mushroom
Research Laboratory. Pant University of Agriculture and Technology. Pantnagar India.
Singh, S., A. Yadav, R.S. Balyan, R. K. Malik and M. Singh, 2004. Control of Ragweed
Parthenium (Parthenium hysterophorus and associated weeds). Weed Techonology 18: 658664.
Smith, J.F. and M.E. Love, 1995. Investigation on to the cultural requirement of a brown
caped Agaricus strain (W4II) isolated from curpressus leaf litter. Journal of Horticultural
Science 70(6): 963-974.
Soto-Cruz, O., G. Saucedo-Castaneda, J.L. Pablos-Hach, M. Gutierrez- Rojas, E. FavelaTirres, 1999. Effect of substrate composition on the mycelial growth of Pleurotus ostreatus.
An analysis by mixture and response surface methodologies. Process Bio chem. 35: 127-133.
Son T.N., 1995. Bioconversion of organic wastes for sustainable Agriculture. Ph.D. Thesis,
Tamil Nadu Agricultural University, Coimbatore.
Srinivas, B., B.N.Gupta, 1997. Chemical composition of wheat. Animal Feed Technology,
65:275–286
Stamets, p., J.S. Chilton, 1983. The mushroom cultivator: a practical guide to growing
mushrooms at home. Agarickon Press, Olympia, wa.
Stamets, P., 2000. Growing gourmet and medicinal mushrooms, Ten Speed Press, Berkeley,
CA., Subramanian, CV., 1995. Mushrooms: Beauty, diversity, relevance. Curr. Sci. 69: 986997.
Steven, K., 2004. Nutritional composition of straws. http://www.Website.Org/humanurc/
chapter 3–4 html. Accessed on 29 Jan 2008.
Stevenson, J.A., P.L. Lentz, 2007. Mushroom. Microsoft® Student 2008 with Encarta
Premium 2008 (DVD). Redmond, WA: Microsoft Corporation.
58
Shukla, P.C., P.M. Talpada and M.B. Pande, 1984. Agro-Industrial byproducts as livestock
feed. Prosopis juliflora pods a new cattle feed source. Technical Bulletin, I.C.A.R. Animal
Nutrition Department, Gujarat Agricultural University, Anand, - 388110, India.
Tekere, M., I. Ncube, J.S. Read, R. Zvauya, 2001 Biodegradation of the organochlorine
pesticide, lindane by a subtropical white rot fungus in batch and packed bed bioreactor
system. EnvironTechnol; 23: 199–206.
Taye Tessema, Rezene Fesseha, Firehun Yirefu, Dereje Tadesse and Tamado Tana, 2009.
Plant protection society of Ethiopia (PPSE). pp. 381-404. In: Abraham Tadesse (eds.).
Increasing crop production through Improved plant production- volume I. PPSE and EIAR,
Addis Ababa, Ethiopia.
Tripathi, D.P., 2005. Mushroom Cultivation. Oxford and IBH Publishing Co. Pvt. Ltd., New
Delhi.
United States Department of Agriculture (USDA), 2003. Mushrooms. Agricultural Statistics
Board. Washington, D.C.
Vats, S.K., R.P. Sood and G.A. Sharoon, 1994. Lantana camara L lignocellulosic substrate
for cultivation of Pleurotus sajor caju. Agriculture and Bioresource Technology 48: 49-52.
Vijayakhader, H.G., 2002. Raga Agricultural University, Rajendranager, Hyderabad, India.
http://www. Edata-center. Com/journals/ 708 ac 68 d 64 b17 c52.
Villa-Cruz, V., G. Huerta-Palacios, J.E. S_anchez-Vazquez, 1999. Fermentation of a mixture
of corn-cobs and coffee pulp for the cultivation of Pleurotus ostreatus. Micol. Neotrop. Appl.
12: 67–74.
Vimal, O.P. and P.D. Tyagi, 1986. Prosopis juliflora Chemistry & Utilization. In: Role of
Prosopis in Wasteland Development. (V.J. Patel, ed.), OPV-1 to 8. Jivrajbhai Patel Agro
forestry Centre, Surendrabag-Kardij - 364 061 (Gujarat).
Wang, D., A. Sakoda, M. Suziki, 2001. Biological efficiency and nutritional value of
Pleurotus ostreatus cultivated on spent beer grain. Biosource Technol 78: 293-300.
Wasser, S.P. and A.L. Weis, 1999. General description of the most important medicinal
higher basidiomycetes mushrooms. International Journal of Medicinal Mushrooms 1: 351370.
Wood, D.A., 1985. Useful biodegradation of lignocellulose. pp. 295-309. In: K.W. Fuller and
J.R., Gallon (eds.). Plant Products and the New Technology, Clarendon, Oxford.
Yamanaka, K., 1997. Production of cultivated edible mushrooms. Food Reviews Intimation
1: 327-333.
Yang, J.H., H.C. Lin and J.L. Mau, 2001. Non-volatile taste components of several
commercial mushrooms. Food Chemistry 72: 465-471.
59
Yildiz, S., U.C. Yildiz, E.D. Gezer, A. Temiz, 2002. Some lignocellulosic wastes used as raw
material in cultivation of the Pleurotus ostreatus culture mushroom. Process Biochem. 38:
301–306.
Zadrazil, F., 1975. Influence of CO2 concentration on the mycelium growth of three Pleurotus
species. European Journal of Applied Microbiology, 1: 327–33.
Zadrazil, F., 1978. Cultivation of Pleurotus. pp 69-88. In “the Biology and cultivation of
Edible mushrooms”. Academic press 21.
Zadrazil, F. and H.C. Dube, 1992. The oyster mushroom: importance and prospects.
Mushroom Re, 1: 25-30.
60
7. Appendix
61
7.1. Appendix I. Analysis of Variance Tables
Appendix table 1. Analysis of variance on days of mycelium formation of Pleurotus species
Source of variation
Degree of freedom
Mean Squares
DMI
Species
2
4.38ns
Substrate
3
66.69**
Species * Substrate
6
0.65ns
Error
24
1.35
**and ns are significant at 0.01 level and non significant, respectively.
Coefficient of Variation: 5.67
DMI= Days of mycelium invasion
Appendix table 2. Analysis of variance on days of pinhead formation of Pleurotus species
Source of variation
Degree of
freedom
Mean Squares
DPF
Species
2
10.33**
Substrate
3
55.35**
Species * Substrate
6
0.55ns
Error
24
0.52
**and ns are significant at 0.01 level and non significant, respectively.
Coefficient of Variation: 2.94
DPF= Days of Pin head formation
Appendix table 3. Analysis of variance on days of fruit body formation of Pleurotus species
Source of variation
Degree of
freedom
Mean Squares
DFF
Species
2
11.73*
Substrate
3
50.85**
Species * Substrate
6
0.71ns
Error
24
1.53
**and ns are significant at 0.01 level and non significant, respectively.
Coefficient of Variation: 4.31
DPF= Days of fruit body formation
62
Appendix table 4. Analysis of variance on Biological efficiency of Pleurotus species
Source of variation
Degree of
freedom
Mean Squares
BE%
Species
2
812.65**
Substrate
3
2032.85**
Species * Substrate
6
102.22**
Error
24
4.79
**and ns are significant at 0.01 level and non significant, respectively.
Coefficient of Variation: 3.88
BE= Biological efficiency
Appendix table 5. Analysis of variance on Production rate of Pleurotus species
Source of variation
Degree of
freedom
Mean Squares
PR%
Species
2
1.58**
Substrate
3
4.03**
Species * Substrate
6
0.18**
Error
24
0.02
**and ns are significant at 0.01 level and non significant, respectively.
Coefficient of Variation: 7.20
BE= Production Rate
Appendix table 6. Analysis of variance on Total yield of Pleurotus species
Source of variation
Degree of
freedom
Mean Squares
TY(g/kg)
Species
2
325058.16**
Substrate
3
813138.24**
Species * Substrate
6
40887.80**
Error
24
1916.35
**and ns are significant at 0.01 level and non significant, respectively.
Coefficient of Variation: 3.88
TY= Total yield
63
Appendix table 7. Analysis of variance on Moisture content of Pleurotus species
Source of variation
Degree of
freedom
Mean Squares
MC%
Species
2
0.79ns
Substrate
3
22.61**
Species * Substrate
6
10.70**
Error
24
0.58
**and ns are significant at 0.01 level and non significant, respectively.
Coefficient of Variation: 0.86
MC= Moisture content
Appendix table 8. Analysis of variance on Dry matter of Pleurotus species
Source of variation
Degree of
freedom
Mean Squares
DM%
Species
2
0.79ns
Substrate
3
22.61**
Species * Substrate
6
10.70**
Error
24
0.58
**and ns are significant at 0.01 level and non significant, respectively.
Coefficient of Variation: 7.04
DM= Dry matter
Appendix table 9. Analysis of variance on Total ash of Pleurotus species
Source of variation
Degree of
freedom
Mean Squares
TA%
Species
2
25.20**
Substrate
3
31.42**
Species * Substrate
6
6.70**
Error
24
0.39
**and ns are significant at 0.01 level and non significant, respectively.
Coefficient of Variation: 6.23
TA= Total ash
64
Appendix table 10. Analysis of variance on Percent organic matter of Pleurotus species
Source of variation
Degree of
freedom
Mean Squares
OM%
Species
2
25.20**
Substrate
3
31.42**
Species * Substrate
6
6.70**
Error
24
0.39
**and ns are significant at 0.01 level and non significant, respectively.
Coefficient of Variation: 0.70
OM= Organic matter
Appendix table 11. Analysis of variance on percent Crude protein of Pleurotus species
Source of variation
Degree of
freedom
Mean Squares
CP%
Species
2
86.09**
Substrate
3
55.85**
Species * Substrate
6
23.18**
Error
24
5.74
**and ns are significant at 0.01 level and non significant, respectively.
Coefficient of Variation: 7.06
CP= Crude protein
Appendix table 12. Analysis of variance on percent crude fiber of Pleurotus species
Source of variation
Degree of
freedom
Mean Squares
CF%
Species
2
16.95**
Substrate
3
26.10**
Species * Substrate
6
3.38**
Error
24
0.86
**and ns are significant at 0.01 level and non significant, respectively.
Coefficient of Variation: 12.16
CF= Crude fiber
65
7.2. Appendix II Chemical Composition of the Substrates
Appendix table 13: Carbon and nitrogen content of substrates
Substrate
Carbon (%)
Nitrogen (%)
C:N
Wheat straw
46.46
0.59
78.75
Lantana camara
51.13
1.54
33.2
Parthenium hysterophorus
50.09
1.5
33.39
Prosopis juliflora
54.12
1.2
45.1
7.3. Appendix III Oyster mushroom cultivation on different substrate
Appendix Figure 1 . Mycelium invasion of oyster mushroom on wheat straw (left) and on
Lantana camara (right)
Appendix Figure 2. Mycelium invasion of oyster mushroom on Parthenium hysterophorus
(left) and on Prosopis juliflora (right)
Appendix Figure 3. Fruit body formation of oyster mushroom on Wheat straw (left) and on
Lantana camara (right)
67
Appendix Figure 4. Fruit body formation of oyster mushroom on Parthenium hysterophorus
(left) and on Prosopis juliflora (right)
68
7.4. Appendix IV Meteorological Data
Appendix Figure 5: Minimum and maximum temperature and relative humidity data during experimental period.