COMPARATIVE STUDY OF THE GROWTH AND YIELD OF THREE CULTIVATED 
Pleurotus SPECIES ON SELECTED TROPICAL TREES SAWDUSTS 
 
 
BY 
 
CALEB ADEWALE OTUNLA 
B.Sc.; M.Sc. (Microbiology, Ibadan), PGDE (Ibadan). 
 
A Thesis in the Department of Botany, 
Submitted to the Faculty of Science 
in partial fulfillment of the requirements for the Degree of 
 
MASTER OF PHILOSOPHY 
 
of the 
 
UNIVERSITY OF IBADAN 
 
 
 
AUGUST, 2015. 
UNIVERSITY OF IBADAN LIBRARY 
ABSTRACT 
The quest to boost dietary protein production from readily available and affordable sources is 
ever increasing in developing countries. The indigenous edible Pleurotus species (oyster 
mushrooms), which grow naturally on wood wastes, are potential protein supplements. However, 
little information is available on the usage of selected tropical trees for optimum mushroom 
cultivation. Therefore, this research was designed to investigate the growth and yield of oyster 
mushrooms on sawdust of selected tropical trees. 
Mature stems of Mangifera indica L. (PTBG0000039360), Senna siamea Lam. 
(BISH0000032830) and Azadirachta indica A. Juss (BISH0000015188) were harvested and 
identity authenticated at the National Horticultural Research Institute (NIHORT), Ibadan. The 
samples were air-dried, separately milled into sawdust, composted and used as three substrates. 
The fourth substrate was the mixed bed derived from the mixtures of the three substrates in ratio 
1:1:1 by weight. Three mushroom species (Pleurotus ostreatus, P. pulmonarius and P. tuber-
regium) were collected from Mycology Unit, NIHORT. A total of 108 polyethylene substrate 
bags (27 for each substrate) were filled with 300 g of the sawdust, each tightly packed, sterilised 
and inoculated with 30 g each of mushroom spawn. Mycelial growth was determined using 
standard method. Fruiting body production was obtained for the three mushrooms on all the 
substrates and sclerotia weight recorded at different Weeks of Composting Intervals (WCI) of 4, 
8 and 12. Biological and production efficiencies were determined using mathematical methods. 
The experiment was a 4 x 3 x 3 factorial arrangement laid out in a complete randomised design 
with three replicates each. The data were analysed using descriptive statistics and ANOVA at p 
=0.05. 
The longest and the shortest mycelial extensions (13.3 and 4.6 cm) were observed in P. 
pulmonarius grown on A. indica at 4WCI and 8WCI respectively. At 12WCI, the highest Fruit 
Weight (FW) of 86.8±1.2 g for P. pulmonarius was observed on S. siamea which was not 
significantly different from P. ostreatus (84.9±1.2 g) on M. indica. The most significant 
Biological Efficiencies (BE) of 82.7% and 80.9% for P. pulmonarius and P. ostreatus at 12WCI 
respectively were observed on S. siamea and M. indica. At 12WCI, the Production Efficiency 
(PE) was highest (42.8%) for P. ostreatus on M. indica and 41.1% for P. pulmonarius on S. 
siamea. Also, the highest mean sclerotia weight of 42.1±0.9 g was obtained for P. tuber-regium 
on M. indica at 12WCI. However, at 4WCI, the least FW (13.7±0.1 g) was in P. pulmonarius 
cultivated on S. siamea while the least BE (13.0%) was observed for P. pulmonarius on S. 
siamea. Also, the least PE value of 4.9% for P. pulmonarius was observed on S. siamea while 
the least sclerotia weight of 9.0±0.6 g was obtained on mixed bed. The longer the decomposition 
period of the substrate, the more significant was the yield. 
The best substrate for the production of fruiting body of Pleurotus ostreatus and sclerotia of 
Pleurotus tuber-regium was Mangifera indica while Senna siamea was most suitable for 
Pleurotus pulmonarius.  
Keywords: Biological and production efficiencies, Composting intervals, Oyster 
                 mushrooms, Tropical trees sawdust 
Word count: 488 
 
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DEDICATION 
This work is dedicated to the Almighty God, the “I AM THAT I AM” (Exodus 3: 14) and the 
Fountain of all knowledge. Surely, your persistence in God’s presence will perpetuate your 
prosperity.                                                       
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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ACKNOWLEDGEMENTS 
 My unreserved and profound gratitude goes to my Supervisor, Prof. S.G. Jonathan, for 
his immense tutelage, tolerance and support in the course of this research study. I really 
appreciate your invaluable encouragement right from the inception of this research study. I 
cannot forget your “push-ups” when it appeared moving forward became a serious challenge. 
You are not just a lecturer, but indeed a loving and caring father who is always interested in the 
progress of other people. It is impossible for you to be stagnant, you will always advance in the 
mighty name of Jesus. God bless you mightily sir. He will continue to promote you (amen). 
 I am quite appreciative of Dr. (Mrs) O.O. Idowu for her unforgettable assistance. Thanks 
for going all out at helping me to “dust” the rusty knowledge I had in the academic line I now 
towed. Sincerely, you have been my physical guiding angel and a “technical adviser”. You will 
not run out of God’s anointing in Jesus name (amen). Likewise, Dr. O.J. Olawuyi, you have been 
so wonderful to me. You have been a friend, a mentor and a God-sent instructor. I pray you will 
always excel both spiritually and physically (amen). Not left out is Dr. A.A. Jayeola and Dr. 
‘Nike Ogunshe. Thank you for your indelible suggestions. God of promotion will always 
promote you (amen). 
 My special appreciation goes to the Executive Director of the National Horticultural 
Research Institute (NIHORT), Dr. (Mrs) A.O. Olufolaji and her other management members for 
the golden opportunity to proceed on this course of study. Not left out is the Head of 
Department, Botany, Prof. A.C. Odebode for your support and concern. I will never forget going 
out of your way, even at odd times, to attend to my needs. The Lord will always be your help 
(amen). 
 I am indeed grateful for all the support and indelible suggestions of Prof. and Mrs 
Gbadegesin, Prof. V.O. Adeyeye, Dr. (Mrs) O.S. Adebayo, Dr. J.A. Akinfasoye, Dr. ‘Layi 
Akintola, and Mrs. H.T. Akinlolu.. Stay lifted. For the right and sound footing morally, 
spiritually and academically, I say a big thank-you to my parents, Mr and Mrs L.A. Otunla. Stay 
blessed.  
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 My special regards goes to Mrs. F.O. Akinrinsola, Mrs. G.A. Majekadegbe and Mr. O. 
Daropale of the mushroom unit, NIHORT, Ibadan for their support and encouragement. The 
efforts of my colleagues at making this dissertation a huge success are worthy of been 
mentioned. I appreciate the constructive criticisms and corrections of Dr. E.O. Ajayi, Dr. Abdul-
Rafiu Monsur, Mr. Tope Modupeola, Mrs. ‘Sola AdeOluwa, Mrs. ‘Dunni Akinpelu, Mrs. ‘Wura 
Ajijola, Mrs. Ogunjinmi, Mr. ‘Niyi Atiba, Mrs. Fajinmi, Mrs. Idowu-Agida, Mrs. E.A. 
Adesegun, Dr. (Mrs) Fagbola, Dr. (Mrs) Badmus, Mr. and Mrs. ‘Gbenga Olanrewaju and Mr. 
Nurudeen Sunmola to mention a few. God bless you all. 
 Highly indebted am I to the members of the PRAYER MILITANTS SQUAD and the 
Full Gospel Business Men’s Fellowship International (FGBMFI), University Area Chapter for 
your support in prayers, cash and kind. God will forever renew your anointing (amen).  
Evangelist ‘Tayo Otunla, you are loved. Not left out are my siblings for their support and 
understanding when the going was tough. God bless you all (amen). 
 Finally to my family; my lovely and ever-loving wife, Otunla Omowumi Elizabeth, you 
are so invaluable. My children; Praise, Abimbola and Glory, you are sources of GREAT JOY. 
You are my darlings. Truly, life is not in duration but in donation and contribution. I have never 
and will never regret you all coming to my life for God has been promoting me from glory to 
glory. I love you all. 
 
 
 
 
 
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CERTIFICATION 
I certify that this work was carried out by Mr. C.A. Otunla in the Department of Botany, 
University of Ibadan. 
 
 
……………………………………………………………. 
Supervisor 
S. G.  Jonathan 
B.Sc, M.Sc. and Ph.D. (Ibadan) 
Professor of Food and Environmental Mycology, Department of Botany 
University of Ibadan, Nigeria. 
 
 
 
 
 
 
 
 
 
 
 
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TABLE OF CONTENTS 
Contents                                                                                                                           Pages 
Title page           i 
Abstract           ii 
Dedication           ii 
Acknowledgements          iv 
Certification           vi 
Table of contents          vii 
List of tables           ix 
List of plates           xi 
List of figures           xii 
CHAPTER ONE: INTRODUCTION 
1.1 Mushrooms          1 
1.2 Oyster mushrooms         1 
1.2.1 Pleurotus ostreatus         2 
1.2.2 P. pulmonarius          2 
1.2.3 P. tuber-regium          2 
1.3 Substrates for mushroom cultivation       3 
1.4 Choice of Oyster mushrooms        3 
1.5 Choice of the selected tropical trees       4 
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1.6 Statement of problem         4 
1.6.1 Justification          4 
1.6.2 Aim           4 
1.6.3 Objectives          5 
CHAPTER TWO: LITERATURE REVIEW 
2.1 Origin and Geographical Distribution of Mushrooms     6 
2.1.1 Botany of Mushrooms         7 
2.1.2 Taxonomy of Mushrooms        7 
2.1.3 Morphology of Mushroom        8 
2.2 Mushroom cultivation         11 
2.3 Nutritional Requirements for the growth of Mushrooms     12 
2.4 Economic importance         13 
2.5 Major phases in mushroom cultivation       15 
2.6 The selected tropical plants as substrate       19 
2.6.1 MANGO PLANT         19 
2.6.2 Local names          19 
2.6.3 Chemical components         20 
2.6.4 Scientific Classification         20 
2.6.5 Mango stem as mushroom substrate       20 
2.7 CASSIA PLANT          20 
2.7.1 Common names          22 
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2.7.2 Scientific classification         22 
2.7.3 Cassia stem as mushroom substrate       23 
2.8 NEEM PLANT          23 
2.8.1 Local names          24 
2.8.2 Scientific classification of neem       24 
2.8.3 Neem as mushroom cultivation material      24 
CHAPTER THREE: MATERIALS AND METHODS 
3.1 Source of spawn (Mushroom seed)       25 
3.1.1 Preparation of mother spawn        25 
3.1.2 Preparation of the planting spawn       25 
3.2 Collection of substrates         25 
3.2.1 Substrate preparation         26 
3.2.2 Substrate composting         26 
3.3 Research Design          26 
3.4 Mycelial ramification         27 
3.4.1 Growth of the mycelia of Pleurotus ostreatus, P. pulmonarius  
        and P. tuber-regium         27 
3.5 Fruiting body production         27 
3.5.1 Cultivation of the fruiting bodies of P. ostreatus, P. pulmonarius   27 
3.6 Cultivation of the sclerotia of P. tuber-regium      28 
3.7 Evaluation of growth and yield characters in Pleurotus ostreatus  
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      and P. pulmonarius         28 
3.7.1 Data analysis          29 
CHAPTER FOUR: RESULTS 
4.1 Preparation of spawn         30 
4.2 Effects of different substrate types on the growth of mushroom mycelium 
      and fruit body production         30 
4.2.1 Effects of different substrate types on the growth of the mycelia of 
        P. ostreatus, P. pulmonarius and P. tuber-regium     30 
4.2.2 Effects of different substrate types on the fruiting bodies production of 
         P. ostreatus and P. pulmonarius       30 
4.3 Effects of different substrate types on the yield of the sclerotia of 
      Pleurotus tuber-regium         66 
CHAPTER FIVE: DISCUSSIONS        71 
CHAPTER SIX: CONCLUSIONS        76 
REFERENCES          77 
 
 
 
 
 
 
 
 
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LIST OF TABLES 
Tables                             Pages 
Table 4.1: Interactions of weeks of composting intervals, substrates, varieties 
                  and their combinations on growth and yield     36 
Table 4.2: The effect of substrate types on the yield and growth parameters of 
                 Pleurotus ostreatus and P. pulmonarius      40 
Table 4.3: The performance of the substrate as affected by weeks of composting intervals 
                 (WCI) on the yield parametersof Pleurotus ostreatus and P. pulmonarius 45 
Table 4.4: The performance of the substrate as affected by weeks of composting  
                  intervals (WCI) on the biological and production efficiencies of 
                  Pleurotus ostreatus and P. pulmonarius      47 
Table 4.5: The performance of the substrates as affected by weeks of composting intervals 
                 (WCI) on the growth parameters of Pleurotus ostreatus and P. pulmonarius 49 
Table 4.6: The mean performance of two mushroom varieties (Pleurotus ostreatus and 
                  P. pulmonarius) as affected by  weeks of composting intervals (WCI) on the 
                 yield parameters         51 
Table 4.7: The mean performance of two mushroom varieties (Pleurotus ostreatus 
                  and P. pulmonarius) as affected by weeks of composting intervals (WCI) on  
                  the growth parameters        54 
Table 4.8: Effect of substrates x varieties interaction on the yield parameters of 
                 Pleurotus ostreatus and P. pulmonarius      55 
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Table 4.9: Effect of substrates x varieties interaction on the biological and production 
                 efficiencies of Pleurotus ostreatus and P. pulmonarius    57 
Table 4.10: Effect of substrates x varieties interaction on the growth parameters of 
                   Pleurotus ostreatus and P. pulmonarius      59 
Table 4.11: Effects of substrates x varieties x weeks of composting intervals (WCI) 
                   interaction on the yield parameters of Pleurotus ostreatus and P. pulmonarius 61 
Table 4.12. Effects of substrates x varieties x weeks of composting intervals (WCI) interaction 
                   on the growth parameters of Pleurotus ostreatus and P. pulmonarius  64 
Table 4.13. Effects of substrates x varieties x weeks of composting intervals (WCI) interaction 
                   on the growth parameters of Pleurotus ostreatus and P. pulmonarius  67 
Table 4.14. The interaction effects of substrate types and weeks of composting intervals 
                   (WCI) on the yield (g) of the sclerotia of Pleurotus tuber-regium  70 
 
 
 
 
 
 
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LIST OF FIGURES 
    Figures                  Pages 
Figure 2.1: Morphology of mushroom fruiting body      9 
Figure 2.2: Life cycle of mushroom        10 
Figure 2.3: Flow Chart of Mushroom Production      16 
Figure  4.1: Combined effect of weeks of composting intervals (WCI)  
                   on the yield  parameters of Pleurotus  ostreatus and P. pulmonarius  37 
Figure 4.2: Combined effect of weeks of composting intervals (WCI) 
                   on the growth parameters of Pleurotus ostreatus and P. pulmonarius  38 
Figure 4.3: The effects of varieties on the yield parameters of  
                   Pleurotus ostreatus and P. pulmonarius      42 
Figure 4.4: The effects of varieties on the growth parameters of 
                  Pleurotus ostreatus and P.   pulmonarius     43 
Figure 4.5: Performance of the substrate as affected by weeks of composting 
                  interval (WCI) on the biological and production efficiencies 
                  of Pleurotus ostreatus and P. pulmonarius     54 
 
            
 
 
 
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LIST OF PLATES 
    Plates                      Pages 
Plate 4.1: Spawn preparation (Sorghum seeds in bottles ready for sterilization) 31 
Plate 4.2: Growth of spawn in bottles one week after inoculation   32 
Plate 4.3: Mushroom spawn fully grown in bottles and ready for use  33 
Plate 4.4: Growth of mycelia on the different substrate types   34 
Plate 4.5: The fruiting bodies of Pleurotus ostreatus on substrates   35 
Plate 4.6: Sclerotia of  P. tuber-regium on the substrates    68 
Plate 4.7: Harvested sclerotia of P. tuber-regium     69 
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CHAPTER ONE 
INTRODUCTION 
1.1 Mushrooms 
The word mushroom is believed to have originated from the french word “mousseron” 
derived from the word moss (Mau et al., 1993). Mushrooms belong to the Kingdom Mycetae 
which are non-green, edible fungi and that they are large heterogeneous group with various 
shapes, sizes and appearance. They are macro fungi with distinctive fruit bodies which are 
visible to the naked eye (Alexopoulos and Mims, 1979). They are neither plants, due to lack of 
chlorophyll, nor animals because they store glycogen (Alexopoulos and Mims, 1979). 
Mushrooms are seasonal and highly perishable crop and contain about 90% moisture. They are 
non-traditional horticultural crops of high quality proteins, high fibre value, vitamins and 
minerals (Narayanasamy et al., 2009). Mushrooms gathered from farmlands, fields and meadows 
were valued as food (Quimio et al., 1990). They could be cultivated on composted substrates, 
chemically treated or untreated wood logs, or picked up in the field during the rainy season 
(Aletor, 1995; Kadiri, 1999; Kadiri, 2002). All edible fungi are saprophytes, and only the 
reproductive structure comes out of the substrate and forms a fruiting body which is visible, 
called mushroom. Although, some mushrooms are unpalatable and others even poisonous, the 
mushrooms of many species are not only edible but also delicious and nutritious (Chang and 
Miles, 1986; Chang and Miles, 1989). 
1.2 Oyster Mushrooms 
Oyster mushrooms are scientifically called Pleurotus and the latin name Pleurotus 
ostreatus means "sideways oyster", referring to the oyster-like shape of the mushroom (Dike et 
al., 2011). Pleurotus species are found to be one of the most efficient lignocelluloses solid state 
decomposing types of white rot fungi (Baysal et al., 2003). Thus, many agricultural and 
industrial wastes can be utilized as substrates for the production of Pleurotus species (Baysal et 
al., 2003). They require shorter growth time when compared to other edible mushrooms, they 
demand few environmental controls, their fruiting bodies are not very often attacked by diseases 
and pests and they can be cultivated in a simple and cheap way (Jwanny et al., 1995; Patrabansh 
and Madan, 1997). Sawdust and sugarcane bagasse were once reported as the best substrates for 
growing of Oyster Mushroom than other agro-based substrates (Ahmed, 1998). 
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1.2.1 Pleurotus ostreatus 
P. ostreatus is an excellent producer of the industrially important enzyme laccase and 
also used for the decolorization of anthraquinone dye (Ho u et al., 2004). Although, it contains 
lower concentration of major elements like phosphorus, potassium and calcium but trace mineral 
elements like magnesium, boron, cadmium, iron and manganese are at a higher concentration 
than that of Agaricus bisporus (Vetter, 1994). It has extensive use in biodegradation (Adamovic 
et al., 1998), immunomodulatory, anti-hypocholesterolemic, anti-mutagenic and anti-tumour 
activities (Wasser, 2002a; Hossain et al., 2003; Lakshmi et al., 2004; Maiti et al., 2011).  
1.2.2 P. pulmonarius 
It is commonly known as the Indian Oyster, Italian Oyster, Phoenix Mushroom, or the 
Lung Oyster, is a mushroom very similar to Pleurotus ostreatus, the pearl oyster, but with a few 
noticeable differences (Stamets, 2000). The caps of this mushroom are much paler and smaller 
than that of P. ostreatus. It also prefers warmer weather than ostreatus and will appear later in 
the summer. Otherwise, the taste and cultivation of the two species is generally described as 
largely the same (Stamets, 2000). It is widespread in temperate and subtropical forests 
throughout the world. In the eastern United States, this species is generally found on hardwoods, 
while in the west it is commonly found on conifers (Stamets, 2000). 
 A polysaccharide called β-D-Glucan from P. pulmonarius reduced sensitivity to pain in 
mice (Baggio et al., 2010), a basis for new analgesic medications (Baggio et al., 2011). While a 
glucan from it showed potent anti-inflammatory and analgesic properties, a methanol extract of it 
displayed anti-inflammatory and antitumor activity (Jose et al., 2002; Smiderle, 2008). The 
extracts of P. pulmonarius may slow the proliferation of cancer cells and be useful as an 
adjuvant to cancer therapies and can halt the progression of diabetes (Badole et al., 2008; 
Wasonga et al., 2008; Lavi et al., 2010). It may be effective in the treatment of hay fever, cause a 
significant reduction in sneezing and nasal rubbing by inhibiting the release of histamine, can be 
used in the treatment of colitis and also inhibit colon cancer formation associated with colitis in 
mice (Yatsuzuka et al., 2007; Lavi et al., 2010; Lavi et al., 2011). Extracts of P. pulmonarius 
have antimicrobial properties (Ramesh and Pattar, 2010). 
1.2.3 P. tuber-regium 
 It is a popular edible mushroom and is considered a profound health promoting 
mushroom in traditional Chinese medicine (Isikhuemhen et al., 2000; Huang, 2002). It has 
nutritive values and some medicinal properties which include relief for stomach ailments, fever, 
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asthma, smallpox, high blood pressure, and cancer (Wong et al., 2011). It is believed by many 
Nigerians to be capable of curing ailments such as headaches, stomach pain, small pox, fever and 
chest pain (Oso, 1977a; Isikhuemhen and Okhuoya, 1995; Okhuoya et al., 1996). It is used in the 
Asaba area of Nigeria in herbal preparation for pregnant women to aid the development of 
foetus. 
  In Ghana, the sclerotia are used mainly for fattening of malnourished babies and as one 
of the ingredients in the embalming of dead bodies (Okhuoya et al., 1998). The fruiting bodies of 
P. tuber-regium is highly nutritive and very rich in protein, while sclerotium is rich in fiber, 
especially non-starch polysaccharides (Kadiri and Fasidi, 1990), mainly composed of bioactive 
β-glucans responsible for pharmacological actions (Cheung and Lee, 2000; Tao et al., 2006). It 
has antihyperglycemic, antihyperlipidemic, and antioxidant properties (Huang et al., 2012). 
Analysis of its sclerotia has shown the presence of Calcium, Magnesium, Iron and Zinc 
(Okhuoya and Ajerio, 1988).  
1.3 Substrates for mushroom cultivation 
 Mushrooms are grown on great variety of substrates. Mushroom substrate may be defined 
as lignocellulotic material which supports the growth, development and fruiting of mushroom 
and their choice depends on availability and cost (Chang and Miles, 1988a).  
 A lot of studies were reported on the various substrates for mushroom production namely 
straws of rice (Oryza sativa), wheat (Triticum vulgare), ragi (Elucine coracana), bazra 
(Pennisetum typhoides), sorghum (Sorghum vulgare), maize (Zea mays), wood of poplar 
(Populus robusta), oak (Quercus luecothricopora), horse chest nut (Aesculus indica), Acasia 
species, chopped banana pseudostem, cotton stalk, pea shells and poplar sawdust (Saidu et al., 
2011). Most of the commercial producers of mushroom in Malaysia are currently using sawdust 
and rice husk (Saidu et al., 2011).  
 Substrates may also be obtained from various plant remnants without enrichments by 
expensive additives. Composted or uncomposted wheat and paddy straw, banana leaves, 
sugarcane bagasses and leaves, wheat bran, rice husk, sawdust etc can be used as substrate for 
growing mushroom (Gupta, 1986). 
1.4 Choice of Oyster mushrooms 
Oyster mushroom is the second most cultivated edible mushroom worldwide after 
Agaricus bisporus (Sánchez, 2010). Members of the mushroom genus Pleurotus form a 
heterogeneous group of edible species of high commercial importance (Georgios et al., 2004). It 
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has economic and ecological values with medicinal properties (Gregori et al., 2007; Sánchez, 
2010; Khan and Tania, 2012). It has abilities to grow over a wide range of temperatures utilizing 
various lignocelluloses (Sánchez, 2010).  
Oyster mushroom cultivation also help in managing organic wastes whose disposal is 
very complicating and time consuming (Das and Mukherjee, 2007). It converts a high percentage 
of the substrate to fruiting bodies thus increasing profitability (Sanchez, 2010). Particularly, P. 
ostreatus requires a shorter growth time in comparison to other edible mushrooms, demands few 
environmental controls, their fruiting bodies are not often attacked by diseases and pests, and 
they can be cultivated in a simple and cheap way (Sánchez, 2010). 
1.5 Choice of the selected tropical trees 
 The selected tropical trees are mango (Mangifera indica), cassia (Senna siamea) and 
neem (Azadirachta indica) trees. They are readily available in the tropics and consequently, our 
environment. They have lignin and cellulose which support growth of mushrooms and are easily 
converted to sawdust during milling for mushroom cultivation. They can be easily colonized and 
degraded by edible mushrooms (Sánchez, 2010). They have medicinal and seasoning or 
flavouring properties (Lose et al., 2000; Kiepe, 2001; Subapriya and Nagini, 2005). 
1.6 Statement of problem 
Malnutrition cases occur in the most people of developing countries due to poverty level 
and their inaffordability in sourcing for animal proteins. Also, there is an increased demand for 
animal protein in these countries as a result of population explosion and proper diet.  
1.6.1 Justification 
Animal protein is beyond the reach of most people in developing countries because most 
of them live below poverty level (World Bank, 1992). Therefore, there is the need to find out an 
alternative source of protein due to malnutrition as a result of population explosion predicated 
upon by an increasing protein gap. More so, FAO recommended edible mushrooms as food that 
can contribute to protein nutrition of developing countries which depend largely on cereals 
(Islam et. al., 2009). Pleurotus species are one of the choice edible mushrooms that can be 
cultivated in the tropics (Quimio et al., 1990). They possess extensive enzyme systems that are 
able to degrade varieties of lignocellulotic materials (Baysal et al., 2003), thereby utilizing them 
for growth. 
 
 
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1.6.2 Aim 
 The aim of this study is to evaluate the growth and yield of Pleurotus ostreatus, P. 
pulmonarius and P. tuber-regium on the sawdust of some selected tropical trees. 
1.6.3 Objectives 
i. To compare the growth and yield of Pleurotus ostreatus, P. pulmonarius and P. tuber-
regium as affected by weeks of compositing interval. 
ii. To select a suitable substrate for the cultivation of Pleurotus ostreatus, P. pulmonarius 
and P. tuber-regium. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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CHAPTER   TWO 
LITERATURE REVIEW 
2.1 Origin and Geographical Distribution of Mushrooms 
Evidence from molecular systematic studies suggests that many mushroom species may 
be quite ancient (Vilgalys and Sun, 1994; Moncalvo et al., 2000). The Chinese and Japanese 
chronicles indicate that the shiitake mushrooms were collected in the wild and given to the 
Emperor as tribute. Throughout the middle ages and the preceeding century, the Greeks and the 
Romans considered mushrooms as special food and ate mushrooms on special occasions while 
some cultures considered all mushrooms as toadstools and poisonous gifts from the devil. At this 
time, mushrooms could only be obtained in autumn and spring (Quimio et al., 1990).  
Fungi are commonly associated with thunderstorms as it was believed in mythology that 
mushrooms were formed by lightening. According to Oso (1977a), a lot of myths surround the 
origin of Nigerian mushrooms. The myth surrounding the origin of P. tuber-regium says that 
“orunmila”, the supreme deity who represent God on earth had several messengers, one of whom 
was “Ifa”, the god of divination. “Ifa” had 16 disciples one of whom was “Ejiogbe” (the god of 
all goodness) who introduced Pleurotus tuber-regium on earth. Termitomyces robustus origin is 
associated with a poor man called “Ogogo”, who after consulting “orunmila” had some rites 
performed for him. These rites resulted in the man producing T. robustus fruiting bodies which 
people liked and eventually made the man wealthy (Oso, 1977b). Termitomyces microcarpus 
origin is associated with a childless Yoruba woman called “Oran” who approached “Orunmila” 
about her childlessness.“Orunmila” prescribed certain rites for her to perform which the woman 
disobediently failed to do. This made her to start having T. microcarpus fruiting bodies instead 
of having children. Human beings became fond of them due to their sweetness so this mushroom 
was named after the woman and is popularly called “Olu Oran” (Oso, 1977b). The popular 
acceptability of T. globulus is linked with a poor woman who after consulting a “Babalawo” was 
asked to procure a T. globulus fruiting body which was divined upon, making it to multiply many 
fold and the woman became extremely rich after selling the fruiting bodies (Oso, 1977b). 
 Mating compatibility studies have demonstrated the existence of discrete intersterility 
groups (biological species) in Pleurotus, many of which are broadly distributed over one or more 
continents (Andersen and Stasovaki, 1992). At least, fifteen intersterility groups have been 
identified, with each group associated with one or more morphological species (Vilgalys et al., 
1996). 
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2.1.1 Botany of Mushrooms 
Mushrooms are familiar to adults and children alike from their most characteristic shape, 
a rounded cap known as pileus on a central stalk called stipe (Turner and Szczawinski, 1992). 
Mushrooms belong to a large, complex group of organisms called fungi, all of which lack 
chlorophyll, the green substance that enables green plants to manufacture their own food through 
photosynthesis (Turner and Szczawinski, 1992). The fruiting body lasts for a few years but the 
mycelium living on the organic materials in the soil may survive for years (Oyetayo and 
Oyetayo, 2008). The mycelium branches and produces enzymes that digest complex 
carbohydrates, lipids and proteins, which are then easily absorbed by the hyphae. The hyphae 
penetrate the substrate until it is time to form fruiting bodies or to start reproduction (Oei, 2003). 
2.1.2 Taxonomy of Mushrooms 
The modern classification has grouped mushrooms into the Kingdom Mycetae. The large 
fleshy mushrooms are found in two classes of fungi (Eumycophyta); the Basidiomycetes and the 
Ascomycetes. Most of the edible mushrooms belong to Ascomycotina and Basidiomycotina 
(Pathak et al., 2003). Some mushrooms, such as truffles and morels are Ascomycetes, while the 
vast majority of the large fungi are Basidiomycetes (Chang, 1981). The size, texture, shape and 
colour of the fruiting body vary according to the types of genera and species of mushroom 
(Alexopoulos and Mims, 1979). 
The fungal class Basidiomycetes is further divided into sub-class Hymenomycetes, whose 
fruiting bodies are exposed to the air and Gasteromycetes, which have unexposed fruiting layers. 
Members of the sub-class Hymenomycetes comprises of four divisions: gill fungi (fungi with 
gills), pore fungi (fungi with tubes or pores), teeth fungi (fungi that are club or coral-like in 
shape) and jelly fungi (fungi that form jelly-like masses) (Kadiri et al., 2003). On the basis of 
spore print colour, the gill fungi are sub-divided into white spore-genera (example include; 
Amanita sp, Lepiota sp, Pleurotus sp, Tricholoma sp, Lentinus sp), pink-spore genera (which 
include Volvariella sp, Enteloma sp), brown-spore genera (which include; Pholiota sp, 
Cortainarius sp), purple-spore genera (which include; Psalliota sp, Panaelous sp) and black-
spore genera (Coprinus sp). The pore fungi consist of the genera Boletus sp, Boletinus sp and 
Polyporus sp. The teeth and jelly-fungi have only one genus each and these are Clavaria and 
Auricularia respectively (Jonathan, 2002; Kadiri et al., 2003). The Gasteromycetes consist of the 
genera Gaester (star-shaped), Lycoperdon (puff-balls) and Calvatia (giant puff-balls). In general, 
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mushrooms are identified on the basis of their spore prints, morphological features and results 
produced by some chemical tests (Kadiri et al., 2003). 
2.1.3 Morphology of Mushroom 
Mushrooms are fleshy, spore-bearing structures which are easily visible. Some have a 
ring or annulus on the upper part of the stalks while others have none. They are variable in form, 
but they have a common function which is to bear the sexual spores. These spores are borne in or 
on specialized hyphal structures (asci or basidia) which make up a fertile layer called a 
hymenium. Most mushrooms bear their hymenia on gills, while there are also some with 
hymenium-lined canals, tubes, or cavities (Chang and Miles, 1989). Like all filamentous fungi, 
the mushroom hyphae can grow over, into, or through the substrate by the extension of hypha. 
This extension occurs at the tips of the hyphae. The older portions of the hyphae are not capable 
of growth, but they have an important role in supporting the growth of the tip and the 
development of fruiting bodies as new protoplasm is formed and the absorbed nutrients are 
transported to the active growing apices (Chang and Miles, 1989). The morphology of a typical 
mushroom is as shown in figure 2.1. 
Mushrooms are only the reproductive part of the organism- the “fruit”; the main part is a 
seldom-seen mass of tiny thread-like growths, or hyphae, called the mycelium. Ever present but 
usually inconspicuous mycelia penetrate soil, bark, and wood (Turner and Szczawinski, 1992). 
Mushrooms make up only a small fraction- about 5,000 to 10,000 of the total number of fungi, 
which includes an estimated 200,000 or more species, the majority being inconspicuous or 
microscopic. Although many fungi are edible or useful to people in some way, many are 
potentially harmful (Turner and Szczawinski, 1992). The basic life cycle is from spore to 
mycelium to fruiting body, which later bears the spores again (Hayes and Hand, 1981). The 
fruiting stage is the formation of visible mushrooms. Hence, mushrooms are actually fruits of the 
fungus (Okhuoya and Okogbo, 1990). The life cycle of mushroom is as shown in figure 2.2. 
 
 
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          Fig. 2.1: Morphology of mushroom fruiting body 
         Source: Stamets (2000) 
 
 
 
 
 
 
 
 
 
 
 
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Fig. 2.2: Life cycle of mushroom 
Source: Stamets (2000) 
 
 
 
 
 
 
 
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2.2 Mushroom cultivation 
Cultivation of saprophytic edible mushrooms may be the only currently economical 
biotechnology for lignocellulose organic waste recycling that combines the production of protein 
rich food with the reduction of environmental pollution (Obodai et al., 2003). The cultivation of 
edible mushrooms is a prime example of how low-value waste produced primarily through the 
activities of the agricultural, forest and food-processing industries can be converted to a higher 
value commodity useful to mankind. It is a profitable agribusiness, particularly edible oyster 
mushroom, with excellent flavour and taste (Shah et al., 2004). It helps in the conversion of 
lignocellulosic waste material into high quality protein food. 
The cultivation of mushrooms needs preparation of substrate and compost; preparation of 
spawn and seeding of the spawn on suitable substrate for mycelial growth and production of 
fruiting bodies (Meera, 2004). The materials that are most widely adopted for mushroom 
cultivation are `lignocellulosic' materials, the major components of which are cellulose, 
hemicellulose and lignin. These three polymeric substances form the bulk of most plant cell 
walls. 
The white button mushroom,  Agaricus bisporus (Lange) Imbach, is the most commonly 
grown mushroom in Turkey, accounting for up to 94.8% of the total mushroom production, and 
productivity of this species is 20-22% of compost fresh weight (Erkal and Aksu, 2000; Erkel, 
2004). It is produced on a composted mixture of various cereal straws (wheat, rye, corn) hay, 
corncobs, distillers’ grain, cottonseed meal, poultry manure and other raw materials (Van 
Griensven, 1988).  
Mushrooms have been recognized as a high potential converter of cheap celluloses into 
valuable protein (Poppe, 2000). There are about 100 species of edible mushrooms all over the 
world. Unfortunately, it is realized that mushrooms did not receive universal acceptance over the 
years since a number of naturally growing mushrooms are poisonous. Now the situation has been 
changed because the cultivated edible mushrooms are totally safe for human consumption. 
Mushroom cultivation fits in very well with sustainable farming and has several advantages: it 
uses agricultural waste products, a high production per surface area can be obtained, after 
picking; the spent substrate is still a good soil conditioner (NPCS, 2011).  
Pleurotus species can produce a broad spectrum of lignocellulolytic enzymes and have 
been grown on different kinds of sawdust, straw and many other agricultural and industrial 
wastes (Hadder et al., 1993). Various works using different substrates like sawdust (Block et al. 
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1958), paddy straw (Bano and Srivastava, 1962), banana pseudo stems (Jaindaik, 1974), 
newspaper (Hashimoto and Takahashi, 1974), wheat straw (Zadrazil, 1974), cotton and sugar 
cane wastes (Chang,1980), maize cobs (Sivaprakasam and Kandaswamy, 1981), hulled cocoa 
shells (Phettipher, 1987) have been done. Waste paper and used cotton have been reported 
(Shakil et al., 2014). 
Presently three mushrooms namely Pleurotus species (Oyster Mushroom), Volvariella 
volvaceae (Straw Mushroom) and Auricularia spp (Ear Mushroom) are under commercial 
cultivation in Bangladesh.  
Most mushrooms performed better during the rainy season (Aletor, 1995). Lentinus 
subnudus and P. tuber-regium had been successfully cultivated on various cellulolytic 
agricultural wastes (Fasidi and Ekuere, 1993; Fasidi and Kadiri, 1993). Similarly, utilization of 
agricultural waste as growing media for the production of mushroom plays a key role in reducing 
the waste and at the same time useful as a bio-fertilizer (Sher et al., 2011). For this reason, it is 
not necessary to process substrates for cultivation of Pleurotus species (Khan and Chaudhary, 
1987; Yalinkiliç et al., 1994). 
 Other basal ingredients that may be used include palm-bunches, straw and corn cobs or 
mixtures thereof. Idowu (2003) observed that oil palm bunch waste singly and in combination 
with other substrates was stimulatory to the growth of the mushroom mycelia which resulted in 
higher fruiting body yield and biological efficiency. Furthermore, it was observed that P. 
pulmonarius performed best on oil palm bunches in comparison with other substrates 
investigated (Idowu, 2003). Mushroom production is completely different from growing green 
plants because it does not contain chlorophyll, and therefore depends on other plant material 
(substrate) for its food. Therefore, the substrate is an important item for growing mushroom.  
2.3 Nutritional Requirements for the growth of Mushrooms 
Sawdust is the most popular basal ingredient in synthetic formulation of substrate used to 
produce shiitake mushrooms (Miller and Jong, 1987). In almost all cases, the efficiency of these 
waste constituting substrates is considerably enhanced when supplemented with protein-rich 
materials such as bran of rice and wheat. Regardless of the main ingredient used, starch-based 
supplements such as wheat bran, rice bran, millet, rye, corn, etc are added to the mixture in a 10 
to 40% ratio (dry weight) to the main ingredient. Royse and Sánchez (2008) indicated that 
mushroom yields may also be stimulated by supplementation of first break mushroom compost 
with hydrolyzed protein, commercial supplements and crystalline amino acids. These 
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supplements serve as nutrients to provide an optimum growing medium (Royse et al., 1990). 
However, all kinds of lignocellulosic substances are likely to be used as substrate for Pleurotus 
species cultivation, the main and co-substrate differ among countries and even regions on the 
basis of availability and cost (Balazs, 1995; Croan, 1999; Labuschagne et al., 2000; Oei, 2003). 
Mushroom cultivation ensures their availability throughout the world irrespective of 
season since cultivated mushrooms can be grown under different climatic conditions on cheap, 
readily available agro wastes. The cultivation process guarantees edibility and serve as the most 
economically viable process for the bioconversion of low value wastes produced primarily from 
the activities of agricultural, forest and food processing industries to produce higher value fungal 
protein for human consumption (Wasser, 2002b). Pleurotus species are also found to be one of 
the most efficient lignocelluloses solid state decomposing types of white rot fungi (Baysal et al., 
2003). Thus, many agricultural and industrial wastes can be utilized as substrates for production 
of Pleurotus species (Zadrazil and Brunnert, 1981; Platt et al., 1983; Platt et al., 1984; Baysal et 
al., 2003). 
All edible fungi are saprophytes (Chang and Miles, 1989) and they need organic matter to 
decompose (Oei, 2003). Mushroom hyphae liberate large amounts of extracellular enzymes 
which bring about the degradation of the many types of macromolecules, such as cellulose, 
hemicelluloses, lignin, protein, etc., present in the substrate. The simple, soluble smaller 
molecules resulting from the activities of these extracellular enzymes are then absorbed by the 
fungal cells. Thus, mushroom hyphae can easily grow and colonize the substrate (Chang and 
o
Miles, 1989). Mushrooms require a temperature of 20-32  Celsius and about 35-90% humidity. 
They also require adequate ventilation, diffused light and semi-darkness. Too much light makes 
mushrooms dark in colour. 
2.4      Economic importance 
 For many reasons, the fungi of the Pleurotus genus have been intensively studied in 
many parts of the world; they have high gastronomic value. They are able to colonize and 
degrade a large variety of lignocellulosic residues, reduce wastes, control environmental 
pollution and the spent mushroom compost (SMC) can be utilised as bio-fertilizer (Sher et al., 
2011). 
Fungi have been extensively studied by Mycologists in educational research fields. 
Antibiotics, therapeutic agents have been produced for medicinal use from some fungi such as 
Penicillium notatum, Aspergillus species, Pleurotus species, Lycoperdom species, Polyporus 
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species (Jonathan, 2002). They have less carbohydrate so they are believed to be suitable for 
diabetic patients. Fresh mushrooms have very limited life and hence they need to be consumed 
within few hours but processing and canning increases their shelf life to few months. Osmotic 
dehydration is one of the important methods of processing mushroom which involves drying 
technology of mushroom. Mushrooms are very popular in most of the developed countries and 
are becoming popular in many developing countries like India (NPCS, 2011). 
Mushrooms are regarded as highly nutritive food delicacies and are important features of 
human diets worldwide (Oso, 1981). Edible mushrooms have been consumed as food and 
delicacies in many cultures (Fasidi and Kadiri, 1990; Jonathan et al., 2008). They are good 
sources of non-starchy carbohydrates, dietary fibre, mineral and vitamins (Bano and 
Rajarathanum, 1988). Several species of edible mushroom have been reported to have protein as 
high as 50.8% dry matter (Eiker, 1993; Alofe et al., 1996), thus surpassing most vegetable 
protein sources. Mushrooms have high contents of qualitatively good protein, crude fibre, 
minerals but are poor sources of lipids (Fasidi and Kadiri, 1990). They have high nutrient value 
of almost twice that of any other vegetable or fruits (Sivrikaya et al., 2002). 
 Mushrooms are superior to many vegetables and beans in their nutritive value. They are 
very rich in protein, vitamins and minerals. Fresh mushrooms contain about 85% water and 3.2% 
protein. But dried mushrooms have low water content and protein level is as high as 34 to 44% 
while the fat content is less than 0.3% (NPCS, 2011). Mushrooms are rich sources of mineral 
elements and vitamins, with mineral elements already detected being potassium, phosphorus, 
calcium, sodium, magnesium, zinc, iron, manganese, nickel, copper, chromium, cobalt and 
vitamins (B, C, K) and niacin (Ogundana and Fagade, 1982, Zakhary et al., 1983). They are rich 
in protein, minerals, and vitamins, and they contain an abundance of essential amino acids 
(Sadler, 2003). Therefore, mushrooms can be a good supplement to cereals (Chang and Buswell, 
1996).  
Mushrooms have been used as human food for centuries, being valued particularly for the 
variety of flavours and textures they can provide (Sadler, 2003). Furthermore, Fasidi and 
Akwakwa (1996) stated that edible mushrooms are eaten in Nigeria as alternatives to meat and 
also for medicinal purposes. Mushrooms can also be canned for consumption and exported to 
foreign countries (Jonathan and Fasidi, 2003a). Protein tends to be present in an easily digestible 
form and on a dry weight basis. The protein content of mushroom normally ranges between 20 
and 40% which is better than many legume sources like soybeans and peanuts, and protein-
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yielding vegetable foods (Chang and Buswell, 1996; Chang and Mshigeni, 2001). Moreover, 
mushroom proteins contain all the essential amino acids needed in the human diet and are 
especially rich in lysine and leucine which are lacking in most staple cereal foods (Chang and 
Buswell, 1996; Sadler, 2003). Mushrooms are low in total fat content and have a high proportion 
of polyunsaturated fatty acids (72 to 85%) relative to total fat content, mainly due to linoleic 
acid. The high content of linoleic acids is one of the reasons why mushrooms are considered a 
health food (Chang and Mshigeni, 2001; Sadler, 2003). Furthermore, they contain significant 
amounts of carbohydrates and fibres (Chang and Buswell, 1996).  
It can be naturally found in tropical and subtropical rainforests, and can be artificially 
cultivated (Maziero et al., 1992). Mushroom is appreciated because of its delicious taste and that 
it has high quantities of proteins, carbohydrates, minerals (calcium, phosphorus, iron) and 
vitamins (thiamin, riboflavin and niacin) as well as low fat (Manzi et al., 1999). Earlier in his 
investigation, Arora (1986) observed that over the centuries, mushrooms, especially the wild 
poisonous forms or toadstools, became objects of fear and distrust because of the stories of 
mushroom poisoning. Most poisoning cases were characterized by extreme pain and suffering 
before death which were recorded not only in America but also in Great Britain. 
2.5 Major phases in mushroom cultivation  
Mushroom farming is a complex business, which requires precision. The major 
practical steps/ segments of mushroom cultivation are: (a) selection of an acceptable mushroom 
species; (b) secreting a good quality fruiting culture; (c) development of robust spawn; (d) 
preparation of selective substrate/ compost; (e) care of mycelial (spawn running); (f) 
management of fruiting/ mushroom development; and (g) harvesting mushrooms carefully 
(Chang 1999). The major phases in mushroom cultivation are as shown in figure 2.3. 
 
 
 
 
 
 
 
 
 
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Selection of an acceptable mushroom species 
 
 
Secreting a good quality fruiting culture 
 
 
 
 
Development of robust spawn 
 
 
 
Preparation of selective substrate/ compost 
 
 
 
Care of mycelial (spawn running) 
 
 
 
Management of fruiting/ mushroom development 
 
 
 
 
Harvesting of mushrooms carefully 
 
                                   Fig. 2.3: Flow Chart of Mushroom Production 
 
 
 
 
 
 
 
 
 
 
 
 
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(a) Selection of Acceptable Mushroom Species/ Strains 
Before any decision to cultivate a particular mushroom is made, it is important to 
determine if that species possess organoleptic qualities acceptable to the indigenous 
population or to the 33 international markets, if suitable substrates for cultivation are 
plentiful, and if environmental requirements for growth and fruiting can be met 
without excessively costly systems of mechanical control (Chang, 2000). 
(b) Secreting a Good Quality Fruiting Culture 
A "fruiting culture" is defined as a culture with the genetic capacity to form fruiting 
bodies under suitable growth conditions. The stock culture which is selected should 
be acceptable in terms of yield, flavour, texture, fruiting time, etc. (Stamets, 2000) 
(c) Development of Robust Spawn 
A medium through which the mycelium of a fruiting culture has grown and which 
serves as the inoculum or "seed" for the substrate in mushroom cultivation is called 
the "mushroom spawn". Failure to achieve a satisfactory harvest may often be traced 
to unsatisfactory spawn used. Ragunathan et al. (1996) reported that consideration 
must also be given to the nature of the spawn substrate since this influences rapidity 
of growth in the spawn medium as well as the rate of mycelia growth and filling of 
the beds following inoculation. 
(d) Preparation of Selective Substrate/ Compost 
The process of substrate preparation is broadly termed “composting”. The final 
product of “composting” is called the “compost” or prepared substrate. While a sterile 
substrate free from all competitive micro-organisms is the ideal medium for 
cultivating edible mushrooms, systems involving such strict hygiene are generally too 
costly and impractical to operate on a large scale. Substrates for cultivating edible 
mushrooms normally require varying degrees of pre-treatment in order to promote 
growth of the mushroom mycelium to the practical exclusion of other micro-
organisms. The substrate must be rich in essential nutrients in forms which are readily 
available to the mushroom, and be free of toxic substances which inhibit growth of 
the spawn. It was observed by Stamets (2000) that moisture contents, pH and good 
gaseous exchange between the substrate and the surrounding environment are 
important physical factors to consider. Mushroom substrate may be simply defined as 
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a lignocellulosic material which supports the growth, development, and fruiting of 
mushroom mycelium (Chang and Miles, 1988a).  
 The process for the preparation of substrates has been the subject of much 
scientific and practical interest over the past two decades. The different types of 
mushrooms require different types or substrate/ compost. Lentinula edodes and 
Pleurotus species are fungi that can grow on wood. The composting conditions 
produce a favourable medium for the development of mushrooms due to the 
development of microbial population that paved the way for the subsequent growth 
and fructification of mushrooms. The large amount of substrate left after the 
mushrooms have been harvested is known as spent compost. It is certainly not 
desirable to leave it without utilization because it can be a possible source of 
pollution. The remains of spent compost consists considerable amount of 
lignocellulosic material in addition to the mushroom mycelia as well as other 
products formed by the metabolic activities of the mycelium. Thus, the spent compost 
is capable of supporting further biological activities. For example, the growth of 
another species of edible mushroom is used as fodder for livestock; as a soil 
conditioner in bio-fertilizer; and also in bioremediation. 
(e) Care of Mycelia (Spawn Running) 
Following composting, the substrate is placed in beds where it is generally 
pasteurized by steam to kill off potential competitive microorganisms. After the 
compost has cooled, the spawn may be broadcast over the bed surface and then 
pressed down firmly against the substrate to ensure good contact, or inserted 2 to 2.5 
cm deep into the substrate. Spawn running is the phase during which mycelium grows 
from the spawn and permeates into the substrate. Good mycelial growth is essential 
for mushroom production and depends on proper maintenance of the beds/ 
innoculated substrate, and also of the mushroom house, in terms of temperature, 
moisture content, humidity and aeration (Quimio, 1988). 
(f) Management of Fruiting/Mushroom Development 
The management of fruiting under suitable environmental conditions may differ from 
those adopted for spawn running. Primordial formation occurs which is then followed 
by the production of fruiting bodies. The appearance of mushrooms normally occurs 
in rhythmic cycles called "flushes"(Jandaik and Goyal, 1995). 
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(g) Harvesting Mushrooms Carefully 
 According to Jandaik and Goyal (1995), harvesting is carried out at different 
maturation stages depending upon the species, consumer preferences and market 
value. 
2.6 The selected tropical plants as substrates 
2.6.1 MANGO PLANT 
This plant (Mangifera indica) belongs to the order Sapindales and the family 
Anacardiaceae. It is a group of tropical trees native to North India, Burma, and Malaya. The 
mango tree is believed to have evolved as a canopy layer or emergent species of the tropical 
rainforest of South and South-east Asia (Kaur et al., 1980). It is found in the wild and have been 
introduced to other warm regions of the world. It is the largest fruit-tree in the world, capable of 
a height of one-hundred feet and an average circumference of 12 to 14 feet, sometimes reaching 
twenty 20. This tree can grow up to 90 feet and have a spread of 120 feet or more (Mukherjee 
and Litz, 2009). It is widely grown in the tropics for its delicious fruit. In the United States of 
America, it is grown outdoors in Southern Florida and the warmest parts of California. Young 
plants can be grown indoors as houseplants. Their foliage is dark green and shiny. When new 
leaves unfold, they have a rich brownish-red color (Mukherjee and Litz, 2009). 
 It is believed that the Portuguese transported the mango from their colonies in India to 
their African colonies, although Purseglove (1972) suggested that it might also have been 
introduced to Africa via Persia and Arabia in the 10th century by Arab traders. The Portuguese 
later introduced the mango into Brazil from their African colonies of Mozambique and Angola. 
On the basis of ancient accounts of travellers and the written historical record, it was believed for 
many years that mango must have originated in India and spread outwards from there to South-
east Asia and thence to the New World and Africa (Mukherjee and Litz, 2009).  The mango is 
cultivated commercially throughout the tropics and in many subtropical areas. The Mangifera 
species, like many other tropical fruit trees, are canopy and emergent trees of the tropical 
rainforest (Kaur et. al., 1980). These trees are widely scattered in the tropical rainforest, flower 
erratically and reproduce from large seeds that deteriorate rapidly. 
2.6.2 Local names:  Yoruba: Mango 
    Hausa: Mangwaro 
    Igbo:  Mangolo 
    Efik:  Mangoro 
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2.6.3 Chemical components 
From the young leaves (172 g/kg), bark (107 g/kg), and from old leaves (94 g/kg), 
mangiferin (a pharmacologically active flavonoid, a natural xanthone C-glycoside) is extracted 
from mango at high concentrations (Barreto et al., 2008). Chlorophyll, carotenes, anthocyanins 
and xanthophylls are all present in the fruit. The skin is generally a mixture of green, red and 
yellow pigments, although fruit colour at maturity is genotype dependent. During ripening, the 
chloroplasts in the peel become chromoplasts, which contain yellow and red pigments 
(Krishnamurthy and Subramanyam, 1970; Akamine and Goo, 1973; Salunkhe and Desai, 1984; 
Mitra and Baldwin, 1997). The pulp carotenoids in ripe fruit also vary with respect to cultivar 
(Mitra and Baldwin, 1997). 
2.6.4 Scientific Classification 
Kingdom: Plantae 
Division: Angiospermae 
(Unranked): Rosids 
Order: Sapindales 
Family: Anacardiaceae 
Genus: Mangifera 
Species: M. indica 
2.6.5 Mango stem as mushroom substrate  
 An enhanced growth of mushroom pileus has been observed on mango sawdust (Veena et 
al., 1998) as Ashrafuzzaman et al. (2009) also observed that mango sawdust gave the largest 
diameter of mushroom pileus. Islam et al. (2009) evaluated the growth and yield of Pleurotus 
flabellatus on seven different type of substrates viz. Mango, Jackfruit, Coconut, Jam, Kadom, 
Mahogony, Shiris sawdust with wheat bran and CaCO3 and obtained the maximum biological 
yield on mango sawdust. Islam et al. (2009) also stated that the cost benefit analysis revealed that 
mango and shiris sawdusts were promising substrates for the growing of Oyster Mushroom 
(Pleurotus flabellatus). 
2.7 CASSIA PLANT 
The genus Cassia is in the family Leguminosae in the major group Angiosperms 
(Flowering plants). Cassia is a genus of Fabaceae in the subfamily Caesalpinioideae. Commonly 
called cassias, "cassia" is also the English name of Cinnamomum aromaticum in the Lauraceae 
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(from which the spice cassia bark is derived), and some other species of Cinnamomum. It is 
commonly called Thailand shower, minjiri, or kassod and has many regional names (F/FRED, 
1994). It is native to South and Southeast Asia and is usually planted as a shade tree in cocoa, 
coffee, and tea plantations (National Academy of Sciences, 1984; Webb et al., 1984).  
Senna siamea, the species used for this study, is a medium sized evergreen tree attaining 
5 m height (F/FRED, 1994). It is a medium size tree up to 15-20 cm tall, with a straight trunk up 
to 30 cm in diameter, bole short, crown usually dense and rounded at first, later becoming 
irregular and spreading with dropping branches, bank grey or light brown, smooth but becoming 
slightly fissured with age (Bernard, 2005). It is an evergreen tree commonly cultivated in fuel 
plantation (Smith, 2009). 
It rarely exceeds 20 m height and 50 cm diameter at breast height (Jensen, 1995). It has a 
dense, evergreen, irregular, spreading crown, a crooked stem, and smooth, grayish bark that is 
slightly fissured longitudinally. Its young branches have fine hairs. The leaves are pinnately 
compound with an even leaf arrangement of 7-10 pairs of ovate-oblong leaflets 7-8 cm long and 
1-2 cm wide. Its flowers are yellow, borne in large terminal panicles that are often 30 cm long. 
The flowering period is long, and flowers may often be found at various seasons (Troup, 1921). 
The fruit is a flat pod 15-25 cm long, thickened at both sutures, containing many seeds 
(Gutteridge, 1997).  
It grows well in many environments, but it grows particularly well in lowland tropics 
having mean annual rainfall of 500-2800 mm (optimum about 1000 mm), mean minimum 
o o
temperature of 20 C, and mean maximum temperature of 31 C. In semiarid environments with 
mean annual rainfall of 500-700 mm, it will grow only where its roots have access to 
groundwater and where the dry season does not exceed 4-6 months. Best growth occurs in deep, 
well drained, rich soils with pH 5.5-7.5. It tolerates well drained lateritic or limestone soils and 
moderately acid soils (pH 5.0). It is susceptible to cold and frost and generally does not grow 
well above 1300 in. It requires full sun (Gutteridge, 1997; Davidson, 1985).  
It is native to South and Southeast Asia, from Thailand and Myanmar to Malaysia, 
India, Sri Lanka, and Bangladesh (Khan and Alam, 1996). It has been cultivated worldwide and 
is naturalized in many locations (Gutteridge, 1997). No significant pest or disease damage has 
been recorded, but minor damage can be caused by the wood rot Ganoderma lucidum (Khan and 
Alam, 1996). Insects that damage seed include Caryedon lineaticollisi, Bruchidius maculatipes, 
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Aspergillus niger and Curvularia pallescens while Phaeolus manihotis occasionally causes 
damage to the root system (Gutteridge, 1997).  
It is effective in managing constipation associated with a number of causes including 
surgery, childbirth and the use of narcotic pain relievers (Hill, 1992). It is used locally as anti-
malaria drugs especially when decocted (the leaves and the bark) (Lose et al., 2000). In 
traditional medicine, the fruit is used to charm away intestinal worms and to prevent convulsion 
in children. The young fruits and leaves are also eaten as vegetables in Thailand. The flowers and 
young fruits are used as curries (Kiepe, 2001). 
  2.7.1 Common names        Amharic: Yeferenji digita   
Creole    : Kasya 
                         Filipino :  Robles 
English  : Black-wood cassia, Bombay blackwood, cassia, iron  
wood, kassod tree, Siamese senna, thai copper pod, Thailand  
shower, yellow cassia.  
It has been cultivated for so long that its exact origin is unknown. It is widely planted 
throughout the tropics and is locally naturalized. Plantations were established in the 1920s in 
Ghana, Nigeria and Sierra Leone, mainly for its quality fuel wood (National Academy of 
Sciences, 1984; Webb et al., 1984). 
2.7.2 Scientific classification 
Kingdom: Plantae 
(Unranked): Angiosperms 
(Unranked): Eudicots 
(Unranked): Rosids 
Order: Fabales 
Family: Fabaceae 
Subfamily: Caesalpinioideae 
Tribe: Cassieae 
Subtribe: Cassinae 
Genus: Senna 
Species: S. siamea 
 
 
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2.7.3 Cassia stem as mushroom substrate 
Das and Mukherjee (2007) observed a longer fruiting time when Pleurotus ostreatus was 
cultivated on dry weed plants; Cassia sophera, Leonotis sp, Sida acuta, Parthenium argentatum, 
Ageratum conyzoides, Tephrosia purpurea and Lantana camara. Furthermore, Das and 
Mukherjee (2007) stated that the protein contents of the fruit bodies obtained from Cassia 
sophera, Parthenium argentatum and Leonotis species were better than cultivating it purely on 
rice straw and as well as the rice straw supplemented with weeds. 
2.8 NEEM PLANT 
The neem tree (Azadirachta indica) originated in India and the word NEEM is derived 
from Sanskrit Nimba which means ‘bestower of good health’ (Chaturvedi et al., 2003). It is a 
medicinal plant belonging to the Meliaceae family and indigenous to Southern Asia (Akula et al., 
2003). The tree (the seeds of which contain up to 45 per cent of a non-edible oil), grows quickly, 
drought resistant and ideal for re-forestation of semi-arid areas.  
It has been used for centuries in Asia as insecticides, fungicides and anti-conceptionals in 
popular medicine. Almost every part of this tree: seeds, leaves, roots, bark, trunk and branches 
have multiple uses (Chaturvedi et al., 2003; Hashmat et al., 2012). It has also been known as 
Ravisambha – sun-ray like effects in providing health. It is recommended for planting in African 
and Asia by many international organizations and regarded as a source of fuel wood in other 
countries (Chaturvedi et al., 2003).  
 The neem tree has been venerated through the ages in the Indian countryside as it 
provided hope in any situation and the faith in the miraculous healing powers of this amazing 
tree led patients with incurable diseases to adopt neem as way of life. It has been extensively 
used in Ayurveda, Unani and Homoeopathic medicine and has become a cynosure of modern 
medicine. It is a valuable source of unique natural products for development of medicines against 
various diseases and also for the development of industrial products (Hashmat et al., 2012). It 
elaborates a vast array of biologically active compounds that are chemically diverse and 
structurally complex. More than 140 compounds have been isolated from different parts of neem. 
 All parts of the neem tree- leaves, flowers, seeds, fruits, roots and bark have been used 
traditionally for the treatment of inflammation, infections, fever, skin diseases and dental 
disorders. Neem leaf and its constituents have been demonstrated to exhibit immunomodulatory, 
anti-inflammatory, antihyperglycaemic, antiulcer, antimalarial, antifungal, antibacterial, 
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antiviral, antioxidant, antimutagenic and anticarcinogenic properties (Biswas et al., 2002; 
Subapriya and Nagini, 2005). 
The tree protects itself from pests with a cocktail of pesticidal compounds called 
limonoids. The most common found in neem include azadirachtin, salannin, meliantriol and 
nimbin. The first is chemically similar to insect hormones that control the process of 
metamorphosis as insects pass from larva to pupa to adult, and appears to block the hormones 
that insects need to moult (Subapriya and Nagini, 2005).      
2.8.1 Local names:   Yoruba: Eke-oyinbo 
     Hausa: Daldejiya/ Dogonyaro 
     Igbo:  Ogwu iba 
2.8.2 Scientific classification of neem 
Kingdom: Plantae 
Order: Rutales 
Family: Meliaceae 
Tribe: Melieae 
Genus: Azadirachta 
Species: A. Indica 
                            Source: Girish and Shankara, 2008 
2.8.3 Neem as mushroom cultivation material 
It has been reported that neem extracts can be used as botanicals to improve and enhance 
mushroom growth and yield. Neem extracts played an important role on the yield and 
productivity of Agaricus bisporus (Polat et al., 2000) and regarded as potential alternatives to 
conventional pesticides for the control of mushroom phorid fly (Erler et al., 2009). Inam-ul-haq 
et al. (2010) and Shah et al. (2011) observed that the extract also reduced the incidence of 
microbes in Oyster mushroom production and is more preferable than chemicals due to their 
lethal effects during human mushroom consumption. 
 
 
 
 
 
 
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CHAPTER THREE 
MATERIALS AND METHODS 
3.1 Source of spawn (Mushroom seed) 
 The spawns of the selected mushrooms; Pleurotus ostreatus, P. pulmonarius and P. 
tuber-regium, were obtained from the mushroom unit, NIHORT, Ibadan. The inocula used were 
generated from the mushroom sporophore obtained at Songhai farm in Porto Novo in Benin 
Republic and maintained on Potato Dextrose Agar (PDA) for regular sub-culturing as described 
by Quimio et al. (1990). 
3.1.1 Preparation of mother spawn 
 Extraneous debris was picked from Sorghum seeds (Sorghum bicolor), soaked in water 
for 2 hrs, drained and boiled for 10 minutes. The seeds were then drained again. About 1% 
calcium carbonate (CaCO3) was added to the seeds to adjust the pH to 7.5. The aliquot portions 
o
of the seeds were poured into 200 ml bottles and sterilized at 121 C for 15 min and allowed to 
 o
cool down to room temperature, 30 ± 2 C. The sterilized substrates (sorghum seeds) in the 
bottles were separately inoculated with actively growing mycelia of all the selected cultivated 
 o
mushrooms. These inoculated substrate bottles were then incubated in the dark room at 30 ± 2 C 
for 15 days. 
3.1.2 Preparation of the planting spawn 
  Sorghum seeds were prepared by removing unwanted particles and soaked overnight. 
They were drained in a strainer the following morning to remove excess water (to about 65% 
moisture content), calcium carbonate (CaCO3) of about 1% was added to the seeds to adjust the 
pH to 7.5 and poured into 200 ml bottles. The mouth of these bottles were plugged with cotton 
o
wool, covered with aluminium foil and sterilized in an autoclave at 121 C for 15 min. They 
 o
were allowed to cool to room temperature of 30 ± 2 C and inoculated asceptically (in an 
inoculating chamber) with freshly prepared mother spawn of all the selected cultivated 
mushrooms separately according to the method described by Quimio et al. (1990). These were 
 o
then incubated under the conditions of 70% relative humidity and at the temperature of 30 ± 2 C 
in a dark room for two weeks. They were kept in the refrigerator until when needed. 
 3.2 Collection of substrates 
The mango (Mangifera indica), Cassia (Senna siamea)  and neem (Azadirachta indica) 
trees used for this research work were obtained within the premises of the National Horticultural 
Research Institute (NIHORT), Ibadan located in the forest Savannah zone of South-West Nigeria 
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o o
(Latitude 7  22`N, Longitude 3  50`E). Their identities were authenticated at the Floriculture 
Unit of NIHORT and assigned voucher numbers as follow: Mangifera indica L. 
(PTBG0000039360), Senna siamea Lam. (BISH0000032830) and Azadirachta indica A. Juss 
(BISH0000015188). The research work was conducted at the Mycology Laboratory in NIHORT. 
3.2.1 Substrate preparation  
The selected tropical trees (substrates) were taken to Sanngo sawmill within Ibadan 
metropolis for milling into sawdust to improve their water retention capacity. The sawdust of 
these tropical trees was sundried for one week and stored to reduce the presence of inhibitory 
substances. Drying and further storage will ensure further decomposition of the sawdust, and 
permit faster growth of the fungi (Chang and Miles, 1988b). 
3.2.2 Substrate composting 
  The sawdusts (substrates) were separately soaked overnight and pressed the following 
morning to remove excess water until the moisture content was about 65% and about 1% 
calcium carbonate (CaCO3) was added to adjust the pH to 7.5. Polyethylene bags of size 25 x 15 
cm were filled with the substrates with each bag weighing 300 g and packed tightly. The neck 
of the bag was made using heat resistant PVC (Poly Vinyl Chloride) tube. The opening was 
covered with a cotton plug through which the spawn was inoculated later. 
3.3 Research Design 
The three levels of composting interval used in this research were 4, 8 and 12 weeks. The 
experiment was a 4 x 3 x 3 factorial arrangement laid out in a complete randomised design with 
three replicates each. For all the experiments, each treatment was replicated three times at each 
of the three levels of composting interval. For the evaluation of mycelia growth and fruiting body 
 o
production at each level, incubation was carried out at the room temperature of 30 ± 2 C.  
A total of 36 substrate bags were produced for each variety of the mushroom. Therefore, 
a sum total of 108 substrate bags were produced for the three varieties of the mushrooms. 
 WEEKS OF COMPOSTING INTERVAL (Weeks) 
SUBSTRATES 4 8 12 
Mango 3 replicates 3 replicates 3 replicates 
Cassia 3 replicates 3 replicates 3 replicates 
Neem 3 replicates 3 replicates 3 replicates 
Mixed Bed 3 replicates 3 replicates 3 replicates 
 
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3.4 Mycelial ramification 
3.4.1   Growth of the mycelia of Pleurotus ostreatus, P. pulmonarius and P. tuber-regium. 
 The sawdusts of mango, cassia, neem and their mixed bed (fourth substrate) were 
evaluated. The fourth substrate was derived from the mixtures of the three substrates in ratio 
1:1:1 by weight. They were separately moistened with water and left overnight. Portions of each 
substrate were packed in a boiling tube of size 16 x 150 mm, plugged with cotton wool, covered 
o
with aluminum foil before sterilization in an autoclave at 121 C for 15 min. These boiling tubes 
were inoculated asceptically with actively growing mycelia of the test mushrooms (P. ostreatus, 
P. pulmonarius and P. tuber-regium) after cooling. Vertical mycelia extension was taken every 
other day from the first day of inoculation for two weeks. 
3.5 Fruiting body production 
3.5.1 Cultivation of the fruiting bodies of P. ostreatus, P. pulmonarius. 
The substrates were separately moistened until the moisture content was about 65% and 
left overnight. The moisture content was considered appropriate if there was no escape of water 
when a handful of the mixture was pressed. Calcium carbonate (1%) was added to adjust the pH. 
Polyethylene bags of size 25 x 15 cm were filled with each substrate, each bag weighing 300 g, 
and tightly packed together. The neck of the bag was prepared by using heat resistant PVC (Poly 
Vinyl Chloride) tube. The opening was covered with a cotton plug and wrapped with aluminum 
foil. The neck served as the opening through which the spawn was introduced. The bags were 
o
sterilized in an autoclave at 121 C for 15 min, and allowed to cool to room temperature.  
 o
 After cooling to room temperature of 30 ± 2 C, each of the bags was inoculated 
separately with 30 g of the inoculum (10% by weight of the substrate in each bag) for two of the 
selected cultivated mushrooms (P. ostreatus and P. pulmonarius) through the neck with three 
replicates per treatment. They were incubated in the dark room for 30 days from the day of 
spawning to allow the mycelia to ramify in the substrates. The fully colonized bags were brought 
out for weighing with an electronic weighning balance and then transferred into the cropping 
house where fruiting bodies emerge and then harvested manually by gently twisting the base of 
each fruiting body without leaving any remnant on the bags to avoid rotting. These procedures 
were carried out on each of the four substrates at the three levels of weeks of composting 
intervals (WCI) of 4, 8 and 12. 
 
 
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3.6 Cultivation of the sclerotia of P. tuber-regium. 
The sawdusts (substrates) of mango, cassia, neem and their mixed bed were prepared. 
About 300 g each of the various sawdust types were packed in polyethylene bags. The neck was 
made with heat resistant polyvinyl chloride (PVC) pipe plugged with cotton wool and covered 
o
with aluminum foil. These bags were also sterilized in an autoclaved at 121 C for 15 min, 
 o
allowed to cool down to ambient temperature of 30 ± 2 C and thereafter inoculated with the 
freshly prepared spawn of P. tuber-regium. The spawned substrate bags were incubated in the 
 o
dark at 30±2 C. Each treatment was replicated three times.  
About 3½ months after spawning, harvesting of the sclerotia was done. Sclerotia were 
harvested from the substrates when the substrate moisture and the mycelium were dried up. 
Substrates clinging to the sclerotia were removed and the sclerotia were weighed. 
3.7 Evaluation of growth and yield characters in Pleurotus ostreatus and P. pulmonarius 
The following data were collected on the mushroom: number of fruiting bodies (NF), fruit 
weight (FW), average fruit weight, width of pileus, length of stipe, primordial initiation and days 
to full colonization, biological and production efficiencies.  
i. Number of Fruiting Bodies: 
Fully opened fruiting bodies of P. ostreatus and P. pulmonarius were harvested per flush by 
gently twisting their base without leaving remnants on the bags and recorded. 
ii. Fruit weight 
The weight of the harvested fruiting bodies of P. ostreatus, P. pulmonarius and the sclerotia 
of P. tuber-regium were taken with a sensitive weighing balance and recorded. 
iii. Average fruit weight 
This was calculated as the ratio of total fruit weight to the total number of fruits. 
Average fruit weight =          Total fruit weight  
                                                      Total number of fruits harvested 
iv. Width of pileus 
With the aid of a slide calliper, the width or diameter of the pileus of P. ostreatus and P. 
pulmonarius were measured and recorded. 
v. Length of stipe 
The length of stipe of P. ostreatus and P. pulmonarius were taken using a ruler. 
 
 
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vi. Primordial initiation 
This was carried out by counting the number of days to first appearance of mushroom 
primordia (pin-head formation). 
vii. Biological efficiency 
This was calculated by using the formula: 
                  Biological efficiency (%) = Total biological yield (g)   X 100 
                     Total substrates used (g)              
viii. Production efficiency 
This was calculated as follow:  
  Production efficiency = Mushroom life weight (g)       X 100 
                                                Substrate weight before cropping (g) 
where; mushroom life weight is the total weight of mushrooms harvested till the nutrients 
in the substrate were expended, while the substrate weight before cropping was the weight 
of the substrate taken just before it was transferred to the cropping house. 
3.7.1 Data analysis 
     The data were analysed using descriptive statistics and ANOVA. Significant means were 
separated using Duncan‘s multiple range test (DMRT) at 5% level of probability (Gomez and 
Gomez, 1984). 
 
 
 
 
 
 
 
 
 
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CHAPTER FOUR 
RESULTS 
4.1 Preparation of spawn 
 The various stages of spawn preparation and growth are shown in plates 4.1- 4.3. 
4.2 Effects of different substrate types on the growth of mushroom mycelium and fruit 
body production. 
4.2.1 Effects of different substrate types on the growth of the mycelia of P. ostreatus, P. 
pulmonarius and P. tuber-regium. 
  It was observed that all the substrates used for this study supported the growth of the 
mycelia of the three tested cultivated mushrooms. Their mycelia grew well on all the tested 
substrates (Plate 4.4).                                             
 4.2.2 Effects of different substrate types on the fruiting bodies production of P. ostreatus 
and P. pulmonarius. 
 There were fruiting bodies yield for the two tested mushrooms on the substrates during 
cultivation (Plate 4.5). The mean square result of analysis of variance (ANOVA) of two 
mushroom varieties of the mushrooms above grown on four substrates is presented in Table 4.1. 
The effect recorded high significant mean squares for all the parameters except the width of 
pileus which was not significant. Likewise, the mean square value for the interaction of 
substrates x varieties was found not to be significant in the number of days for full mycelia 
colonization and days to primordia initiation. Significant means were observed for the three-way 
interaction effect of substrates x varieties x weeks of composting intervals (WCI) except days for 
full mycelia colonization that was significant at p < 0.05 (Table 4.1). 
 Figure 4.1 represents the effect of weeks of composting intervals (WCI) on the yield 
parameters of Pleurotus ostreatus and P. pulmonarius. As WCI increases, the yield parameters 
also increased. 
 Figure 4.2 represents the effect of weeks of composting intervals (WCI) on the growth 
parameters of Pleurotus ostreatus and P. pulmonarius on the biological efficiency (BE), 
production efficiency (PE), mycelia extension, days to full mycelia colonization and extension 
per day. While there was a progressive increase in PE and BE, the number of days for full 
mycelia colonization revealed a progressive decrease in values with the highest as 24.83 days at 
4WCI and the least as 20.13 days at 12WCI. Similar trend was observed in the number of days to 
primordial initiation. 
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Plate 4.1: Spawn preparation (Sorghum seeds in bottles ready for sterilization) 
 
 
 
 
 
 
 
 
 
 
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Plate 4.2: Growth of spawn in bottles one week after inoculation 
 
 
 
 
 
 
 
 
 
 
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Plate 4.3: Mushroom spawn fully grown in bottles and ready for use 
 
 
 
 
 
 
 
 
 
 
 
 
 
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B 
A 
   C D 
   
    
    Plate 4.4: Growth of mycelia on the different substrate types 
    A: Mycelial growth on mango (Mangifera indica) sawdust  
    B: Mycelia growth on cassia (Senna siamea) sawdust 
    C: Mycelial growth on neem (Azadirachta indica) sawdust 
    D: Mycelial growth on the mixed bed   
 
 
 
 
 
  
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Plate 4.5: The fruiting bodies of Pleurotus ostreatus on substrates 
 
 
 
 
 
 
 
 
 
 
 
 
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Table 4.1: Interactions of weeks of composting intervals, substrates, varieties and their combinations on growth and yield 
     of Pleurotus ostreatus and P. pulmonarius. 
Source of DF Number Fruit Average Width Length Biological Production Mycelial Full Primordial Extension 
variation of fruits weight  fruit of of stipe  Efficiency  Efficiency  extension mycelial initiation  per day  
weight  pileus  colonization 
** ** ** ** ** ** ** ** ** ** 
 2 662.35 15399.59 11.14 8.68 5.38 13755.44 3975.65 220.08 138.29 307.04 0.15
** 
** ** **  ** ** ** ** ** ** ** 
Substrate (S) 3 7.71 577.39 9.95 0.03 0.60 576.44 127.51 1.92 10.87 12.76 0.05
** ** ** ** ** ** ** ** ** ** ** 
Varieties (V) 1 435.31 404.29 79.91 16.72 24.04 323.30 120.67 14.37 37.56 25.68 0.02
** ** ** ** ** ** ** ** ** ** ** 
S x WCI 6 6.27 95.12 2.51 1.56 0.41 91.32 25.07 10.45 2.44 5.23 0.00
** ** ** ** ** ** ** ** ** ** ** 
V x WCI 2 115.79 43.76 12.46 2.16 2.50 45.83 13.70 25.68 4.26 4.68 0.03
** ** ** ** ** ** ** ** ** 
S x V 3 5.46 385.41 7.66 1.03 0.77 367.07 107.03 5.11 0.74 1.16 0.02
** ** ** ** ** ** ** ** * ** ** 
S x V x WCI 6 9.79 194.09 3.42 1.58 1.54 162.63 39.44 2.02 1.89 3.94 0.00
Error 48 1.18 4.36 0.33 0.04 0.04 2.03 1.33 0.32 0.71 0.75 0.00 
Total 71            
**  Significant at 1% probability 
*    Significant at 5% probability 
WCI: Weeks of Composting Intervals 
 
 
 
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(a) Number of fruits                                        (b) Fruit weight (g) 
      
© Average fruit weight (g)                                    (d) Width of pileus (cm) 
  
(e) Length of stipe (cm)                                          
   
Fig. 4.1: Combined effect of weeks of composting intervals (WCI) on the yield parameters of 
Pleurotus  ostreatus and P. pulmonarius. 
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(a) Biological Efficiency (%)                 (b) Production Efficiency (%) 
 
    © Mycelial extension (cm)                      (d) Full mycelia colonization (days) 
 
(e) Primordial initiation (days)                    (f) Extension per day (days) 
 
Fig. 4.2: Combined effect of weeks of composting intervals (WCI) on the growth parameters of 
Pleurotus ostreatus and P. pulmonarius. 
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The effect of substrate types on the yield of Pleurotus ostreatus and P. pulmonarius is 
represented in Table 4.2. Mango, as a substrate, produced the highest number of fruits (10.17) 
followed by mixed bed and neem which were not significantly different from each other (i.e. 
9.72 and 9.56 respectively). Cassia produced the least (8.61). The heaviest fruit weight was 
observed on mixed bed (52.50 g) followed by cassia (50.45  g) and mango (50.23g) which were 
not significantly different from each other, while the lowest was produced by neem (39.92 g). 
Cassia recorded the highest average fruit weight (6.55 g) but was not significantly different from 
mixed bed, while mango produced the least (5.08 g) which was also comparable to what was 
produced by neem (5.18 g). The longest width of pileus was observed on neem (5.73 cm). 
However, this was not significantly different from all other substrates. 
The longest length of stipe was obtained from the mixed bed (5.21 cm) which was 
comparable to what was obtained from cassia and mango but the least was from neem (4.79 cm). 
Similar trend was observed in both biological and production efficiencies. The fastest mycelia 
extension was observed in neem (9.73 cm) which was not significantly different from that of the 
mixed bed (9.59 cm). The least was observed on cassia (9.04 cm) but was not significantly 
different from mango (9.18 cm). Cassia recorded the longest number of days for full mycelia 
colonization (23.50 days) followed by the mixed bed (22.61 days), as a substrate, while the 
significantly lowest number of days was observed in mango (21.72 days). The longest number of 
days for mushroom primordial initiation was observed in cassia (27.28 days) followed by neem 
(26.22 days). The least was observed on the mixed bed (25.44 days) which was not significantly 
different from mango (25.56 days). The fastest mycelia extension per day was observed on 
mango (0.66 cm) followed by cassia (0.64 cm) and the mixed bed (0.57 cm) while the least was 
on neem (0.55 cm). 
Figure 4.3 showed the effects of varieties on the yield parameters of P. ostreatus and P. 
pulmonarius. In terms of number of fruits, P. ostreatus was more (11.97) than P. pulmonarius 
(7.06). The same trend was observed in the fruit weight. In contrast, the average fruit weight of 
P. pulmonarius (6.82 g) was more than that of P. ostreatus (4.71 g). This trend was also 
observed in the width of pileus and length of stipe. 
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Table 4.2: The effect of substrate types on the yield and growth parameters of Pleurotus ostreatus and P. pulmonarius. 
Substrate Number of Fruit Average Width Length Biological Production Mycelial Full Primordial Extension 
fruits weight fruit of of Efficiency Efficiency extension mycelial initiation per day 
(g) weight pileus stipe (%) (%) (cm) colonization (days) (cm) 
(g) (cm) (cm) (days) 
Mango 10.17a 50.23b 5.08b 5.72a 5.12a 47.84b 23.05a 9.18b 21.72c 25.56c 0.66a 
Cassia 8.61b 50.45b 6.55a 5.64a 5.13a 48.05b 22.66a 9.04b 23.50a 27.28a 0.64b 
Neem 9.56a 39.92c 5.18b 5.73a 4.79b 37.50c 17.50b 9.73a 23.17ab 26.22b 0.55d 
Mixed Bed 9.72a 52.50a 6.25a 5.71a 5.21a 50.03a 22.71a 9.59a 22.61b 25.44c 0.57c 
 
Means with the same letter along the column are not significantly different from one another at p ≤ 0.05
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 Figure 4.4 represents the effects of varieties on the growth parameters of P. ostreatus and 
P. pulmonarius. Higher biological and production efficiencies were observed in P. ostreatus than 
in P. pulmonarius. Number of days for full mycelia colonization observed in P. ostreatus was 
more than that of P. pulmonarius. However, total mycelial extension, number of days for 
primordial initiation and average extension per day were more in P. pulmonarius than in P. 
ostreatus. 
Tables 4.3- 4.5 showed the performance of the substrates as affected by the number of 
weeks of composting interval (WCI). It was generally observed that all the parameters were 
significantly highest at 12WCI except mycelia extension, days to full mycelia colonization and 
primordial initiation. 
Table 4.3 showed the performance of the substrates as affected by the number of weeks 
of composting interval (WCI) on the number of fruits, fruit weight, average fruit weight, width of 
pileus and length of stipe of Pleurotus ostreatus and P pulmonarius. At 4WCI, the highest 
number of fruits (NF) were observed in mango (5.00) followed by mixed bed and cassia which 
were not significantly different from each other (3.83 and 2.83) while the least number of fruits 
was observed in neem (2.67). At 8WCI, the mixed bed produced the highest NF (12.17) followed 
by mango and cassia substrates (11.17 each) with the least from neem (11.00). Neem and mango 
produced the highest NF (15.00 and 14.33 respectively) at 12WCI followed by the mixed bed 
(13.17) while the least was observed in cassia (11.83). The heaviest fruiting body weight (FW) 
was observed in the mixed bed (27.75 g) at 4WCI which was not significantly different from 
what was obtained in mango (25.50 g). This was followed by comparable results in cassia and 
neem (18.68 g and 16.75 g respectively). At 8WCI, mixed bed (56.47 g) produced the heaviest 
FW followed by cassia (52.79 g) which was not significantly different from what was produced 
by mango (50.49 g). The smallest FW was obtained in neem (39.88 g) at 8WCI. Cassia substrate 
(79.90 g) produced the heaviest FW at 12WCI followed by comparable results from mango and 
mixed bed (74.70 g and 73.28 g respectively). The least was produced by neem (63.12 g).  
 
 
 
 
 
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(a) Number of fruits                                           (b) Fruit weight (g) 
  
© Average fruit weight (g)                                       (d) Width of pileus (cm) 
  
(e) Length of stipe (cm) 
 
P ost: Pleurotus ostreatus    P pul: Pleurotus pulmonarius 
Fig. 4.3: The effects of varieties on the yield parameters of Pleurotus ostreatus and P. 
pulmonarius 
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(a) Biological Efficiency (%)   (b) Production Efficiency (%) 
 
     © Mycelial extension (cm)   (d) Full mycelia colonization (days) 
 
    (e) Primordial initiation (days)                           (f) Extension per day (days) 
 
P ost: Pleurotus ostreatus    P pul: Pleurotus pulmonarius 
Fig. 4.4: The effects of varieties on the growth parameters of Pleurotus ostreatus and P.   
pulmonarius 
 
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  The highest average fruit weight, at 4WCI, was obtained in mixed bed (7.31g) which was 
comparable with what was obtained in neem (6.81g). This was followed by cassia (6.68 g) while 
mango substrate produced the lowest (5.31 g). The largest width of pileus obtained at 4WCI in 
cassia and mango were not significantly different from each other (5.47 cm and 5.40 cm 
respectively). This was also followed by comparable results from neem and mixed bed (5.17 cm 
and 5.13cm respectively). At 8WCI, the widest pileus was produced by cassia (5.87 cm) 
followed by neem (5.65 cm) which was significantly different from mixed bed as substrate (5.23 
cm). The shortest was obtained in mango (4.93 cm). Mango (6.83 cm) produced the largest width 
of pileus which was not significantly different from what was obtained in mixed bed (6.77 cm). 
This was followed by neem (6.37 cm) while least was observed in cassia (5.60 cm). In terms of 
length of stipe, the longest was produced on the mixed bed (4.83 cm) at 4WCI but this was not 
significantly different from what was obtained in mango (4.68 cm) and cassia (4.65 cm). The 
least was observed in neem (4.23 cm). At 8WCI, the longest was obtained on cassia (5.40 cm) 
comparable to what was produced by the mixed bed (5.22 cm) as substrates. This was followed 
by similar results from mango and neem (4.77 cm each). At 12WCI, the longest length of stipe 
was produced by mango (5.90 cm) followed by the mixed bed (5.57 cm) comparable with what 
was obtained in neem (5.38 cm). Cassia sawdust produced the least (5.33 cm).  
 Table 4.4 showed the performance of the substrate as affected by weeks of composting 
intervals (WCI) on the biological and production efficiencies of Pleurotus ostreatus and P 
pulmonarius. The biological efficiency (BE) was highest in mixed bed (26.51%) at 4WCI 
followed by mango and cassia (24.28% and 17.79% respectively). The least BE was observed in 
neem (15.95%). Mixed bed (53.78%) was the most biologically efficient substrate at 8WCI. This 
was followed by cassia and mango which were not significantly different from each other. Neem 
possessed the least BE of 37.84% at 8WCI. In contrast, cassia (76.10%) was the most 
biologically efficient substrate at 12WCI followed by mango (71.14%) comparable to the mixed 
bed (69.79%). The least was also observed in neem (58.69%) similar to what was observed in 
other WCI. The production efficiency (PE), as observed, in the mixed bed (10.77%) was the 
greatest at 4WCI followed by mango (9.66%). This was followed by comparable results from 
cassia and neem (7.14% and 6.54% respectively). The same trend was observed at 8WCI.
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Table 4.3: The performance of the substrate as affected by weeks of composting intervals (WCI) on the yield parameters 
               of Pleurotus ostreatus and P. pulmonarius. 
Substrate Number of fruits Fruit weight (g) Average fruit weight (g) Width of pileus (cm) Length of stipe (cm) 
x WCI 
 Weeks Weeks Weeks Weeks Weeks 
 4 8 12 4 8 12 4 8 12 4 8 12 4 8 12 
Mango 5.00a 11.17ab 14.33a 25.50a 50.49bc 74.70b 5.31c 4.72b 5.21bc 5.40a 4.93d 6.83a 4.68a 4.77b 5.90a 
Cassia 2.83b 11.17ab 11.83c 18.68b 52.79b 79.90a 6.68b 6.08a 6.88a 5.47a 5.87a 5.60c 4.65ab 5.40a 5.33c 
Neem 2.67bc 11.00b 15.00a 16.75b 39.88c 63.12c 6.81ab 4.24b 4.51c 5.17b 5.65b 6.37b 4.23b 4.77b 5.38bc 
Mixed bed 3.83b 12.17a 13.17b 27.75a 56.47a 73.28bc 7.31a 5.82ab 5.63b 5.13b 5.23c 6.77a 4.83a 5.22ab 5.57b 
 
Means with the same letter along the column are not significantly different from one another at p ≤ 0.05. 
WCI: Weeks of composting intervals
 45 
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However, at 12WCI, PE was greatest in cassia (37.65%) which was not significantly 
different from what was obtained in mango (36.91%). Also, the least was observed in neem 
(28.93%). 
Table 4.5 showed the performance of the substrate as affected by weeks of storage 
compositing interval (WCI) on the mycelia extension, days to full mycelia colonization, days to 
mushroom primordial initiation and extension per day of Pleurotus ostreatus and P. 
pulmonarius. The longest mycelia extension was observed in neem (13.07 cm) at 4WCI followed 
by followed by comparable results from mixed bed and mango (11.38 cm and 11.36 cm 
respectively). The least was obtained in cassia (9.32 cm). Neem (6.44 cm) also produced the 
longest mycelia extension at 8WCI which, however, was not significantly different from what 
was obtained in mango (5.82 cm). A comparable result was obtained in cassia (5.69 cm) while 
the least was produced by the mixed bed (5.63cm). At 12WCI, cassia (12.12 cm) produced the 
longest mycelia extension which was not significantly different from what was obtained in the 
mixed bed (11.75 cm). However, comparable results were observed in mango and neem (10.38 
cm and 9.69 cm respectively) (Table 4.5). 
In terms of full mycelia colonization, at 4WCI, the longest number of days was observed 
in neem (25.67 days) followed by comparable results from other substrates in the order cassia> 
mixed bed> mango (25.17 days, 24.50 days and 24.00 days respectively). At 12WCI, the longest 
was observed on cassia (21.67 days) which was not significantly different from what was 
observed in neem (20.67 days). This was followed by comparable results from mixed bed and 
mango (19.50 days and 18.67 days respectively). 
The primordial was initiated at the shortest number of days, at 4WCI, in mango (28.17 
days) which was not significantly different from what was observed in the mixed bed (28.67 
days) and comparable to cassia (29.67 days). However, the longest was obtained in neem (30.00 
days). All the substrates produced comparable results at 8WCI (Table 5) while at 12WCI, the 
shortest number of days for the mushroom primordial was observed in the mixed bed (20.67 
days). This was followed by comparable results from mango and neem (21.83 days and 21.50 
days respectively). The longest was observed in cassia (24.67 days).  
The longest average extension per day was observed in mango (0.58 cm) at 4WCI 
followed by neem (0.55 cm) which was also significantly different from the mixed bed (0.55 cm)  
 
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Table 4.4: The performance of the substrate as affected by weeks of composting intervals (WCI) 
on the biological and production efficiencies of Pleurotus ostreatus and P. pulmonarius. 
Substrate x Biological Efficiency (%)       Production Efficiency (%) 
WCI 
 Weeks Weeks 
     4     8    12    4     8    12 
Mango 24.28b 40.08c 71.14b 9.66b 22.58b 36.91a 
Cassia 17.79c 50.28b 76.10a 7.14c 23.17ab 37.65a 
Neem 15.95d 37.84d 58.69c 6.54c 17.01c 28.93c 
Mixed Bed 26.51a 53.78a 69.79bc 10.77a 23.79a 33.59b 
 
Means with the same letter along the column are not significantly different from one another at p 
≤ 0.05. 
WCI: Weeks of composting intervals 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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UNIVERSITY OF IBADAN LIBRARY 
while the least was obtained in cassia (0.48 cm). At 8WCI, the same trend was observed (Table 
5). Furthermore, mango substrate (0.73 cm) produced the longest but not stastistically different 
from what was obtained in neem (0.73 cm) at 12WCI. This was followed by the mixed bed (0.65 
cm) which was significantly different from what was obtained in cassia (0.63 cm), as the least 
average extension per day. 
Table 4.6 showed the mean performance of two mushroom varieties as affected by the 
number of weeks of composting intervals (WCI) on the number of fruits, fruit weight, average 
fruit weight, width of pileus and length of stipe. Of the two mushroom varieties, Pleurotus 
ostreatus produced more number of fruits and fruit weights than P. pulmonarius at the three 
levels of the WCI while in terms of average fruit weight, width of pileus and length of stipe, P. 
pulmonarius was more than P. ostreatus.  
At 4WCI, the number of fruits produced by P. osreatus was 4.17, which is statistically 
higher than what was obtained in P. pulmonarius (3.00). P. ostreatus produced 16.25 fruits while 
P. pulmonarius produced 6.50 at 8WCI. At 12WCI, while P. ostreatus produced 15.50 fruits, P. 
pulmonarius produced 11.67. At 4WCI, the total fruit weight (FW) obtained in P. osreatus was 
23.73 g while that of P. pulmonarius was 20.16 g. The total FW observed in P. osreatus at 8WCI 
was 53.83 g while 45.98 g was observed in P. pulmonarius. Furthermore, the FW observed in P. 
osreatus and P. pulmonarius were significantly different from each other (74.37 g and 71.13 g 
respectively).  
 The average fruit weight observed at 4WCI, in P. pulmonarius was more than what was 
obtained in P. ostreatus 7.16 g and 5.90 g respectively). This trend was also observed at 4 and 
8WCI (Table 6). At 8WCI, P. pulmonarius produced an average fruit weight of 7.10g which was 
significantly different from what was obtained in P. ostreatus (3.33 g). At 12WCI, 6.02 g of P. 
pulmonarius was observed which was significantly different from what was observed in P. 
ostreatus (4.19 g). The width of pileus, as observed in P. pulmonarius at 4WCI, was 6.03 cm 
which was significantly different from what was observed in P. ostreatus (4.56 cm). At 8WCI 
also, the width of pileus observed in P. pulmonarius (5.98 cm) was significantly different from 
what was observed in P. ostreatus (4.86 cm) and at 12WCI, the larger was also observed in P. 
pulmonarius (6.54 cm) followed by P. ostreatus (6.24 cm). The longer length of stipe was 
recorded in P. pulmonarius (4.94 cm) followed by what was obtained in P. ostreatus (4.26 cm) at 
4WCI. At 8WCI, it was also longer in P. pulmonarius than in P. ostreatus (5.48 cm and 4.59 cm
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Table 4.5: The performance of the substrates as affected by weeks of composting intervals (WCI) on the growth parameters 
               of Pleurotus ostreatus and P. pulmonarius. 
Substrate x Mycelia Extension (cm) Full Mycelia Colonization Primordial Initiation (days) Extension per Day (cm) 
WCI (days) 
 Weeks Weeks Weeks Weeks 
      4     8     12      4      8     12     4      8    12    4    8   12 
Mango 11.36b 5.82a 10.38b 24.00b 22.50b 18.67b 28.17b 26.67a 21.83b 0.58a 0.68a 0.73a 
Cassia 9.32c 5.69ab 12.12a 25.17ab 23.67a 21.67a 29.67ab 27.50a 24.67a 0.48d 0.56d 0.63c 
Neem 13.07a 6.44a 9.69b 25.67a 23.17ab 20.67a 30.00a 27.17a 21.50bc 0.55b 0.65b 0.73a 
Mixed Bed 11.38b 5.63b 11.75a 24.50b 23.83a 19.50b 28.67b 27.00a 20.67c 0.50c 0.58c 0.65b 
 
Means with the same letter along the column are not significantly different from one another at p ≤ 0.05. 
WCI: Weeks of composting intervals
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UNIVERSITY OF IBADAN LIBRARY 
 respectively). At 12WCI while 6.49cm was observed for P. pulmonarius, what was obtained as 
the length of stipe for P. ostreatus was 4.60 cm. 
 Fig. 4.5 represents the mean performance of two mushroom varieties as affected by the 
number of weeks of composting intervals (WCI) on the biological and production efficiencies of 
Pleurotus ostreatus and P. pulmonarius. Both Biological Efficiency (BE) and Production 
Efficiency (PE) followed the same trend. At 12WCI, P. ostreatus had the best BE (70.11%) 
followed by P. pulmonarius also at the same WCI (67.74%). However, the least was observed in 
P. pulmonarius at 4WCI (19.67%). Furthermore, the greatest PE was recorded in P. ostreatus at 
12WCI (35.23%). This was followed by P. pulmonarius at the same WCI (33.31%). The least 
was also recorded in P. pulmonarius at 4WCI (7.76%). 
 Table 4.7 represents the mean performance of two mushroom varieties as affected by 
weeks of composting intervals period (WCI) on the mycelia extension, days to full mycelia 
colonization, days to mushroom primordial initiation and extension per day of Pleurotus 
ostreatus and P. pulmonarius. The longer mycelia extension (12.65cm) was obtained at 4WCI in 
P. pulmonarius which was significantly different from what was observed in P. ostreatus 
(9.91cm). In contrast, P. ostreatus (6.65cm) produced the longer mycelia extension which was 
significantly different from what was obtained in P. ostreatus at 8WCI while at 12WCI, P. 
pulmonarius produced a longer extension than what was observed in P. ostreatus (11.63cm and 
10.34cm respectively). 
 The shortest number of days observed for the full mycelia colonization was in P. 
ostreatus (18.92 days) at 12WCI while 21.33 days was observed for P. pulmonarius. At 8WCI, a 
shorter number of days (22.83 days) were also observed in P. ostreatus as 23.75 days was 
observed for P. pumonarius. Shorter number of days (24.33 days) was observed for P. ostreatus 
as 25.33 days was recorded for P. pulmonarius at 4WCI (Table 4.7). At 12WCI, the number of 
days for primordial initiation was shorter in P. ostreatus than in P. pulmonarius (21.50 days and 
22.83 days respectively). The same trend was observed at 8WCI while there was no significant 
difference between the two mushroom varieties at 4WCI; 29.00 days for P. ostreatus and 29.50 
days for P. pulmonarius. In terms of average extension per day, it was longer in P. pulmonarius 
than in P. ostreatus (0.58 cm and 0.47 cm respectively). Similar trend was observed at 8WCI, 
0.62cm was recorded for P. pulmonarius while 0.61 cm was recorded for P. ostreatus. However, 
   
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Table 4.6: The mean performance of two mushroom varieties (Pleurotus ostreatus and P. pulmonarius) as affected by 
               weeks of composting intervals (WCI) on the yield parameters. 
Varieties Number of fruits Fruit weight (g) Average fruit weight Width of pileus (cm) Length of stipe (cm) 
x WCI (g) 
Weeks     4      8     12     4     8    12    4    8   12    4    8   12    4    8    12 
P ost 4.17a 16.25a 15.50a 23.73a 53.83a 74.37a 5.90b 3.33b 4.91b 4.56b 4.86b 6.24b 4.26b 4.59b 4.60b 
P pul 3.00b   6.50b 11.67b 20.61b 45.98b 71.13b 7.16a 7.10a 6.20a 6.03a 5.98a 6.54a 4.94a 5.48a 6.49a 
 
Means with the same letter along the column are not significantly different from one another at p ≤ 0.05. 
P ost: Pleurotus ostreatus 
P pul: Pleurotus pulmonarius 
WCI: Weeks of composting intervals 
 
 
 
 
 
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(a) Biological Efficiency (%) 
 
(b) Production Efficiency (%) 
 
P ost: Pleurotus ostreatus    P pul: Pleurotus pulmonarius 
Fig. 4.5: Performance of the substrate as affected by weeks of composting interval (WCI) on the 
biological and production efficiencies of Pleurotus ostreatus and P. pulmonarius. 
 
 
 
 
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UNIVERSITY OF IBADAN LIBRARY 
However, at 12WCI it was longer in P. ostreatus (0.70 cm) than in P. pulmonarius (0.67 cm). 
Table 4.8 represents the effects of substrates x varieties interaction on the number of 
fruits, fruit weight, average fruit weight, width of pileus and length of stipe of Pleurotus 
ostreatus and P pulmonarius. The highest number of fruits (NF) observed during this study was 
in P. ostreatus as produced by neem (12.78) though not significantly different from what was 
obtained in mango (12.22) and also comparable to what was obtained in the mixed bed (11.78). 
The least was observed in by cassia (11.11). Comparable results were obtained on P. 
pulmonarius grown on mango and mixed bed (8.11 and 7.67 respectively) followed by 6.33 and 
6.11 NF from neem and cassia respectively which were not significantly different from each 
other. P. ostreatus grown on mango produced the most significant fruit weight (59.52 g) 
followed by the mixed (52.37 g) which was comparable to what was obtained in cassia (51.04 g). 
The least was recorded in neem (39.66 g).  
The highest weight in P. pulmonarius was recorded on the mixed bed (52.63 g) which 
was significantly different from what was observed in cassia (49.87 g). This was followed by 
comparable results from mango and neem (40.94 g and 40.18 g respectively). The average fruit 
weight observed in mixed bed (5.30 g) with respect to P. ostreatus production was highest but 
this was comparable to what was obtained in cassia (5.19 g). The least was also recorded in neem 
(3.48 g). The most significant average fruit weight was recorded in P. pulmonarius grown on 
cassia (7.91 g). This was followed by comparable results from mixed bed and neem (7.12 g and 
6.89 g respectively). The least was obtained from P. pulmonarius cultivated on mango (5.27 g).  
As observed, the largest width of pileus was recorded in P. ostreatus grown on mixed bed 
(5.47 cm) comparable to what was obtained in mango (5.34 cm). This was followed by the one 
grown on cassia (5.14 cm) which was significantly different from what was observed on neem 
(4.92 cm), as the least. In contrast, neem (6.53 cm) produced the largest width of pileus in terms 
of P. pulmonarius production followed by comparable results from cassia, mango and mixed bed 
(6.14 cm, 6.10 cm and 5.96 cm respectively). The longest length of stipe, in P. ostreatus, was 
produced by the mixed bed (4.87 cm) followed by mango (4.46 cm). Cassia and neem produced 
comparable results (4.31 cm and 4.30 cm respectively). The length of stipe observed in all the 
substrates with respect to P. pulmonarius production was significantly different from each other. 
It was in the order cassia> mango> mixed bed> neem (Table 4.8).  
 
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Table 4.7: The mean performance of two mushroom varieties (Pleurotus ostreatus and P. pulmonarius) 
               as affected by weeks of composting intervals (WCI) on the growth parameters.  
Varieties x Mycelia Extension (cm) Full Mycelia Colonization   Primordial Initiation (days) Extension per Day (cm) 
WCI (days) 
Weeks     4      8     12     4     8    12    4    8   12    4    8   12 
P. ost 9.91b 6.65a 10.34b 24.33b 22.83b 18.92b 29.00a 26.08b 21.50b 0.47b 0.61b 0.70a 
P. pul 12.65a 5.22b 11.63a 25.33a 23.75a 21.33a 29.25a 28.08a 22.83a 0.58a 0.62a 0.67b 
 
Means with the same letter along the column are not significantly different from one another at p ≤ 0.05. 
P ost: Pleurotus ostreatus 
P pul: Pleurotus pulmonarius 
WCI: Weeks of composting intervals 
 54 
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Table 4.8: Effect of substrates x varieties interaction on the yield parameters of Pleurotus ostreatus and P. pulmonarius 
Substrate x Number of fruits     Fruit weight (g) Average fruit weight (g) Width of pileus (cm) Length of stipe (cm) 
Varieties 
 P.  ost P.  pul P.  ost P.  pul P.  ost P.  pul P.  ost P.  pul P.  ost P.  pul 
Mango 12.22a 8.11a 59.52a 40.94c 4.89ab 5.27c 5.34ab 6.10b 4.46b 5.78b 
Cassia 11.11b 6.11b 51.04b 49.87b 5.19a 7.91a 5.14b 6.14b 4.31c 5.94a 
Neem 12.78a 6.33b 39.66c 40.18c 3.48b 6.89bc 4.92c 6.53a 4.30c 5.29d 
Mixed Bed 11.78ab 7.67a 52.37b 52.63a 5.30a 7.21b 5.47a 5.96b 4.87a 5.54c 
 
Means with the same letter along the column are not significantly different from one another at p ≤ 0.05. 
P. ost: Pleurotus ostreatus 
P. pul: Pleurotus pulmonarius
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Table 4.9 showed the effects of substrates x varieties interaction on the biological 
efficiency (BE) and production efficiency (PE) of Pleurotus ostreatus and P. pulmonarius. In P. 
ostreatus, the greatest BE was observed on mango (56.68%) followed by mixed bed (49.87%) 
comparable to what was obtained on cassia (48.61%). The least was recorded on neem (36.72%). 
However, in P. pulmonarius, the greatest BE was recorded in the mixed bed (50.18%) followed 
by cassia (47.50%). Both mango and neem produced comparable results (38.99% and 38.26% 
respectively). The greatest PE recorded for P. ostreatus was on mango (27.99%) followed by 
comparable results from cassia and mixed bed (22.94% and 22.86% respectively). The least was 
observed in neem also (17.30%). Mixed bed (22.57%) recorded the highest PE for P. 
pulmonarius which, however, was not significantly different from what was observed on cassia 
(22.37%). This was followed by comparable results from mango and neem (18.11% and 17.69% 
respectively). 
Table 4.10 revealed the effect of substrates x varieties interaction on the mycelia 
extension, days to full mycelia colonization, days to mushroom primordial initiation and 
extension per day of Pleurotus ostreatus and P. pulmonarius. The longest mycelia extension in 
P. ostreatus was observed in neem (10.04 cm) followed by mixed bed (8.96 cm) which was 
comparable to that of cassia (8.50 cm). The least was recorded in mango (8.25 cm). In P. 
pulmonarius, the longest was obtained on mixed bed (10.21 cm) though not significantly 
different from the one grown on mango (10.11 cm) and comparable to what was recorded in 
cassia (9.58 cm). The least was found in neem (9.42 cm). P. ostreatus recorded the shortest 
number of days to full mycelia colonization in mango (20.89 days) followed by comparable 
results from mixed bed, neem and cassia (22.11 days, 22.22 days and 22.89 days respectively). 
The shortest number of days for full mycelia colonization in P. pulmonarius was observed in 
mango but comparable to that of mixed bed (22.56 days and 23.11 days respectively). This was 
followed by comparable results from cassia and neem (24.11 days each). 
For primordial initiation, the shortest number of days observed for P. ostreatus was in 
mango (24.78 days) but not significantly different from what was obtained in the mixed bed 
(25.22 days). This was followed by neem (25.55 days) but comparable to what was recorded in 
cassia (26.56 days). The mixed bed (25.67 days) however, recorded the shortest number of days 
for primordial initiation in P. pulmonarius followed by mango (26.33 days) which was not 
 
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Table 4.9: Effect of substrates x varieties interaction on the biological and production 
efficiencies of Pleurotus ostreatus and P. pulmonarius. 
Substrates x Varieties Biological Efficiency (%) Production Efficiency (%) 
 P. ost P. pul P. ost P. pul 
Mango 56.68a 38.99c 27.99a 18.11b 
Cassia 48.61bc 47.50b 22.94b 22.37a 
Neem 36.72c 38.26c 17.30c 17.69b 
Mixed Bed 49.87b 50.18a 22.86b 22.57a 
 
Means with the same letter along the column are not significantly different from one another (p 
≤0.05). 
P. ost: Pleurotus ostreatus 
P. pul: Pleurotus pulmonarius 
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UNIVERSITY OF IBADAN LIBRARY 
significantly different from what was recorded for neem (26.89 days). Of all the substrates, 
cassia recorded the longest (28.00 days). The longest average extension per day was observed for 
P. ostreatus grown on mango (0.68 cm) followed by neem (0.60 cm). This was also significantly 
different from what was observed in cassia (0.56 cm) while the shortest was observed in mixed 
bed (0.54 cm). In P. pulmonarius, the longest average extension per day was recorded in neem 
(0.69 cm) followed by mango (0.64 cm). This was also significantly different from what was 
observed in mixed bed (0.61 cm) while the shortest was recorded in cassia (0.55 cm). 
 Table 4.11 represents the effects of substrates x varieties x week of composting interval 
on the number of fruits, fruit weight, average fruit weight and width of pileus of Pleurotus 
ostreatus and P. pulmonarius. At 4 weeks of composting interval (WCI), the most significant 
number of fruits (NF) was observed in P. ostreatus grown on mango (6.00) followed by P. 
pulmonarius grown on mango and mixed bed (4.00 each) though not significantly different from 
P. ostreatus grown on cassia, mixed bed and neem (3.67,3.67 and 3.33 respectively). The least 
NF was however obtained from P. pulmonarius grown on cassia and neem at 4WCI (2.00 each). 
Mixed bed (17.67) produced the highest NF of P. ostreatus though not significantly different 
from what was obtained on cassia (16.67) and comparable to that of mango and neem (15.33 
each) at 8WCI. This was followed by results obtained from P. pulmonarius grown on mango, 
neem and mixed bed (7.00, 6.67 and 6.67 respectively) while the least was observed in P. 
pulmonarius cultivated on cassia (5.56). At 12WCI, P. ostreatus grown on neem (19.67) 
produced the highest NF followed by the result on mango (15.33). The least was also observed in 
P. pulmonarius grown on neem (10.33). 
As observed, P. pulmonarius produced by the mixed bed (28.76±1.55 g) was the heaviest 
at 4WCI though not significantly different from what was obtained in P. ostreatus cultivated on 
mango (28.59±1.32 g). Furthermore, this was comparable to what was observed in P. ostreatus 
grown on mixed bed (26.74±1.44 g). P. pulmonarius was least produced on cassia (13.70±0.14 
g). At 8WCI, the highest fruit weight (FW) was recorded on P. ostreatus grown on mango 
(65.02±1.83 g) followed by P. ostreatus cultivated on mixed bed (58.82±0.58 g) comparable to 
what was observed in P. ostreatus grown on cassia (56.44±1.65 g). P. pulmonarius produced by 
mango and P. ostreatus observed on neem (35.96±2.26 g and 35.05±0.76 g respectively) were 
the least at 8WCI. At 12WCI, P. pulmonarius produced on cassia (86.79±1.22 g) was the highest 
fruit weight (FW) comparable to the result of P. ostreatus grown on mango (84.94±1.23 g). This  
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Table 4.10: Effect of substrates x varieties interaction on the growth parameters of Pleurotus 
ostreatus and P. pulmonarius 
 
Substrates x Mycelia Extension Full Mycelia Primordial Extension per 
Varieties (cm) Colonization  Initiation (days) Day (cm) 
(days) 
 P. ost P. pul P. ost P. pul P. ost P. pul P. ost P. pul 
Mango 8.25c 10.11a 20.89b 22.56b 24.78b 26.33b 0.68a 0.64b 
Cassia  8.50bc 9.58ab 22.89a 24.11a 26.56a 28.00a 0.56c 0.55d 
Neem 10.04a 9.42c 22.22a 24.11a 25.55a 26.89b 0.60b 0.69a 
Mixed Bed 8.96b 10.21a 22.11ab 23.11b 25.22b 25.67c 0.54d 0.61c 
 
Means with the same letter along the column are not significantly different from one another (p 
≤0.05). 
P. ost: Pleurotus ostreatus 
P. pul: Pleurotus pulmonarius 
 
 
 
 
 
 
 
 
 
 
 
 
 
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was followed by P. pulmonarius grown on mixed bed (75.02±1.46 g). The least was recorded P. 
pulmonarius cultivated on neem (58.24±2.11 g). 
The most significant average fruit weight (FW), at 4WCI, was observed in P. 
pulmonarius grown on neem (8.79±0.14 g). This was followed by P. ostreatus produced on 
mixed bed (7.39±0.87 g) with comparable results from P. pulmonarius grown on mixed bed and 
cassia (7.23±0.33 g and 6.85±0.07 g respectively). P. ostreatus produced on mango and neem 
were the least (4.86±0.84 g and 4.82±0.40 g respectively). At 8WCI, P. pulmonarius grown on 
cassia produced the heaviest average FW (8.74±0.96 g) though not significantly different from 
what was observed in P. pulmonarius cultivated on mixed bed (8.30±1.11 g). This was followed 
by P. pulmonarius grown on neem (6.18±0.82 g) while the least was observed in P. ostreatus 
cultivated on neem (2.30±0.22 g). At 12WCI, the heaviest average FW was recorded in P. 
pulmonarius grown on cassia (8.14±0.35 g) followed by P. pulmonarius grown on mixed bed 
and neem with P. ostreatus grown on caasia (6.09±0.39 g, 5.70±0.61 g and 5.63±0.47 g 
respectively). The least was observed in P. ostreatus cultivated on neem (3.31±0.08 g). 
The largest widths of pileus, at 4WCI, were recorded in P. pulmonarius grown on cassia 
and neem (6.40 cm and 6.30 cm respectively). This was followed by P. pulmonarius grown on 
mixed bed (5.97 cm) while the least was observed in P. ostreatus cultivated on neem (4.03 cm). 
At 8WCI, the largest width of pileus was recorded in P. pulmonarius grown on neem (6.87 cm) 
but comparable to what was observed in the same mushroom cultivated on cassia (6.60 cm). This 
was followed by P. pulmonarius grown on mango and P. ostreatus cultivated on mixed bed (5.27 
cm and 5.27 cm respectively). The least was observed in P. ostreatus grown on neem (4.43 cm). 
P. pulmonarius grown on mango (7.60 cm) recorded the largest width of pileus at 12WCI 
followed by P. ostreatus grown on mixed bed (6.83 cm) which was not significantly from P. 
pulmonarius produced by the same mixed bed (6.70 cm). However, the least was observed in P. 
pulmonarius cultivated on cassia (5.43 cm). 
 Table 4.12 showed the effects of substrates x varieties x weeks of composting intervals 
(WCI) interaction on the length of stipe, biological efficiency, production efficiency and mycelia 
extension of Pleurotus ostreatus and P. pulmonarius. As observed, the longest length of stipe, at 
4WCI, was produced by P. pulmonarius grown on cassia (5.17 cm). However, this was not 
statistically different from what was recorded by the same mushroom cultivated on mixed bed 
and mango (5.17 cm and 5.03 cm respectively). This was followed by P. ostreatus grown on
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Table 4.11: Effects of substrates x varieties x weeks of composting intervals (WCI) interaction on the yield parameters of Pleurotus 
ostreatus and P. pulmonarius. 
 
 Substrate Number of fruits  Fruit weight (g) Average fruit weight (g) Width of pileus (cm) 
 WCI (Weeks)    4     8    12     4     8    12     4    8    12    4    8    12 
 Mango 6.00a 15.33ab 15.33b   28.59a 65.02a 84.94ab 4.86d 4.26c 5.54bc 5.37c 4.60cd 6.07d 
P. ost Cassia 3.67b 16.67a 13.00c   23.6lb 56.44bc 73.01bcd 6.51bc 3.42cd 5.63b 4.53d 5.13bc 5.77de 
 Neem 3.33b 15.33ab 19.67a  15.92cd 35.05f 68.02e 4.82d 2.30e 3.31d 4.03e 4.43cde 6.30cd 
 Mixed Bed 3.67b 17.67a 14.00bc  26.74ab  58.82b 71.53d 7.39b 3.33cd 5.17bc 4.30de 5.27b 6.83b 
              
 Mango 4.00b 7.00c 13.33c   22.40b 35.96f 64.46f 5.76cd 5.18c 4.87c 5.43c 5.27b 7.60a 
P. pul Cassia 2.00c 5.56d 10.67d   13.70d 49.13d 86.79a 6.85b 8.74a 8.14a 6.40a 6.60ab 5.43ef 
 Neem 2.00c 6.67c 10.33de   17.58c 44.71e 58.24g 8.79a 6.18b 5.70b 6.30a 6.87a 6.43c 
 Mixed Bed 4.00b 6.67c 12.33cd   28.76a 54.11c 75.02bc 7.23b 8.30a 6.09b 5.97b 5.20bc 6.70bc 
 
Means with the same letter along the column are not significantly different from one another at p ≤ 0.05. 
P ost: Pleurotus ostreatus 
P pul: Pleurotus pulmonarius 
WCI: Weeks of composting interval 
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mixed bed (4.50 cm) and P. pulmonarius cultivated on neem (4.40 cm). The least was recorded 
in P. ostreatus grown on cassia and neem (4.13 cm and 4.07 cm respectively). At 8WCI, P. 
pulmonarius grown on cassia (6.13 cm) also recorded the longest length of stipe but comparable 
with P. pulmonarius produced on neem (5.83 cm). This was followed by P. ostreatus cultivated 
on mixed bed (4.80 cm) while the least was recorded in P. ostreatus grown on neem (3.70 cm). 
P. pulmonarius grown on mango (7.47 cm) was observed to have the longest length of stipe at 
12WCI followed by the same variety cultivated on cassia (6.53 cm). The least was P. ostreatus 
produced on cassia (4.13 cm).  
The highest Biuological Efficiency (BE) observed at 4WCI was found in P. pulmonarius 
cultivated on mixed bed (27.55%) which, however, was comparable to what was recorded in P. 
ostreatus grown on mango and mixed bed (27.23% and 25.47% respectively). They were 
followed by comparable results of P. ostreatus grown on cassia and P. pulmonarius grown on 
mango (22.53% and 21.33% respectively). The least was observed in P. pulmonarius produced 
on cassia (13.04%). P. ostreatus grown on mango (61.92%) was the highest BE at 8WCI. This 
was followed by the same mushroom grown on mixed bed (56.02%). This was comparable to 
what was recorded on the same variety cultivated on cassia (53.77%) while the least was 
observed in P. ostreatus grown on neem (33.09%). At 12WCI, BE was highest in P. pulmonarius 
produced by cassia (82.66%) but comparable to P. ostreatus grown on mango (80.89%). This 
was followed by P. pulmonarius cultivated on mixed bed which was also comparable to what 
was recorded on P. ostreatus grown on cassia (71.45% and 69.53% respectively). The least was, 
however, observed in P. pulmonarius cultivated on neem (55.47%). 
At 4WCI, P. ostreatus grown on mango (11.36%) was the recorded highest Production 
Efficiency (PE). However, this was comparable to the results of P. pulmonarius cultivated on 
mixed bed and P. ostreatus produced by the mixed bed also (11.13 and 10.42% respectively). 
This was followed by P. ostreatus observed on cassia (9.40%) while the least was observed in P. 
pulmonarius grown on cassia (4.89%). Mango (29.77%), as substrate, produced the highest PE at 
8WCI with P. ostreatus. This was followed by the same variety producing comparable results on 
cassia and mixed bed (25.20% and 24.87% respectively). The least comparable PE values were 
obtained in P. pulmonarius grown on mango and P. ostreatus cultivated on neem (15.40% and 
15.36% respectively). At 12WCI, P. ostreatus cultivated on mango (42.84%) also produced the 
highest PE. This was followed by comparable results of P. ostreatus grown on cassia with both 
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P. pulmonarius and P. ostreatus cultivated on mixed bed (34.23%, 33.87% and 33.29% 
respectively).  
At 4WCI, comparable values of the longest mycelial extension was recorded in P. 
pulmonarius grown on neem, mixed bed and mango with P. ostresatus produced by neem (13.25 
cm, 13.13 cm, 13.08 cm and 12.88 cm respectively). This was followed by P. pulmonarius 
cultivated on cassia (11.13 cm) with the least produced by P. ostreatus also grown on cassia 
(7.50 cm). P. ostreatus, at 8WCI, recorded the longest mycelia extension on neem (8.25 cm) 
while all others showed no significant difference. At 12WCI, P. pulmonarius grown on mixed 
bed (12.25 cm) produced the longest mycelial extension but comparable to the same mushroom 
cultivated on cassia, P. ostreatus grown on cassia also with P. pulmonarius produced by mango 
(12.13 cm, 12.11 cm and 11.75 cm respectively). This was followed by comparable results of P. 
ostreatus grown on mixed bed (11.25 cm) and P. pulmonarius on neem (10.38 cm). The least 
comparable results were recorded in P. ostreatus cultivated on mango and neem (9.00 cm each).  
Table 4.13 showed the effects of substrates x varieties x weeks of composting intervals 
(WCI) interaction on the number of days for full mycelia colonization, days to mushroom 
primordial initiation and extension per day of Pleurotus ostreatus and P. pulmonarius. At 4WCI, 
the longest number of days for full mycelial colonization was recorded in P. pulmonarius grown 
on neem (26.00days). However, this was not significantly different from the results obtained in 
P. pulmonarius grown on cassia and mixed bed and P. ostreatus grown on neem and cassia also 
(25.67 days, 25.33 days, 25.33 days, 24.67 days and 24.67 days respectively). This was followed 
by comparable results from P. ostreatus grown on mango (23.67 days) but not significantly 
different from P. pulmonarius grown on mango and P. ostreatus cultivated on mixed bed (24.33 
days and 23.67 days respectively). 
 Also, at 8WCI, the longest was recorded in P. pulmonarius grown on neem (24.67 days). 
This was not significantly different from P. ostreatus cultivated on both mixed bed and cassia 
(24.00 days, 23.67 days, 23.67 days and 23.67 days respectively). This was followed by 
comparable result from P. pulmonarius cultivated on mango (23.00 days) but the least number of 
days for full mycelial colonization was observed in P. ostreatus grown on neem and mango 
(21.67 days and 22.00 days respectively). The longest number of days for full mycelial 
colonization at 12WCI was observed in P. pulmonarius grown on cassia (23.00 days) which
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Table 4.12. Effects of substrates x varieties x weeks of composting intervals (WCI) interaction on the growth parameters of  
                 Pleurotus ostreatus and P. pulmonarius. 
 
 Substrate       Length of stipe (cm)    Biological Efficiency (%)  Production Efficiency (%) Mycelia Extension (cm) 
 WCI(Weeks)    4     8    12     4     8    12     4    8    12    4    8    12 
 Mango 4.33bc 4.70de 4.33fg 27.23a 61.92a 80.89ab 11.36a 29.77a 42.84a   9.63c 6.13b   9.00c 
P ost Cassia 4.13c 4.67de 4.13fgh 22.53bc 53.77bc 69.53cd   9.40ab 25.20b 34.23b   7.50d 5.88b 12.11a 
 Neem 4.07c 3.70f 5.13de 15.16ef 33.09fg 61.92f   6.01cd 15.36e 30.55c 12.88a 8.25a   9.00c 
 Mixed Bed 4.50b 5.30bc 4.80ef 25.47ab 56.02b 68.12de 10.42a 24.87b 33.29b   9.63c 6.00b 11.25b 
              
 Mango 5.03a 4.83d 7.47a 21.33bcd 34.25f 61.39f   7.95bc 15.40e 30.98c 13.08a 5.50b 11.75a 
P pul Cassia 5.17a 6.13a 6.53b 13.04fg 46.79d 82.66a   4.89d 21.14c 41.08a 11.13b 5.50b 12.13a 
 Neem 4.40b 5.83ab 5.63cd 16.74e 42.58e 55.47g   7.08c 18.66d 27.32d 13.25a 4.63b 10.38b 
 Mixed Bed 5.17a 5.13bcd 6.33bc 27.55a 51.53c 71.45c 11.13a 22.71c 33.87b 13.13a 5.25b 12.25a 
 
Means with the same letter along the column are not significantly different from one another at p ≤ 0.05. 
P ost: Pleurotus ostreatus 
P pul: Pleurotus pulmonarius 
WCI: Weeks of composting interval 
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however, was not significantly different from the same variety cultivated on neem (21.67 days). 
But this was comparable to the results from P. ostreatus grown on cassia with P. pulmonarius 
cultivated on mango and mixed bed (20.33 days each). This was followed by P. ostreatus grown 
on neem (19.67 days) while the least number of days was recorded in P. ostreatus cultivated on 
mixed bed (18.67 days). 
The longest number of days for primordial initiation was observed in P. ostreatus grown 
on neem (30.33 days) at 4WCI but comparable with what was obtained in P. pulmonarius grown 
on cassia, neem and mixed bed with P. ostreatus grown on cassia (30.00 days, 29.67 days, 29.00 
days and 29.33 days respectively). The least was however recorded in P. ostreatus grown on 
mixed bed (23.33 days). At 8WCI, the longest was observed in P. pulmonarius cultivated on 
neem but not significantly different from that of cassia (29.33 days and 28.00 days respectively). 
Comparable results were recorded in P. pulmonarius grown on mixed bed and mango with P. 
ostreatus cultivated on cassia (27.67 days, 27.33 days and 27.00 days respectively). The least 
number of days was, however, observed in P. ostreatus grown on neem (25.00 days) but not 
significantly different from the results of P. ostreatus cultivated on mixed bed and mango (26.33 
days and 26.00 days respectively).  
The least number of days were generally observed at 12WCI when compared with the 
other weeks of composting intervals. At 12WCI, P. pulmonarius grown on cassia (26.00 days) 
produced its primordial at the longest number of days. This was followed by comparable results 
from P. ostreatus cultivated on cassia and P. pulmonarius grown on mango (23.33 days each). 
The least was observed in P. ostreatus cultivated on mango (20.33 days) but not significantly 
different from P. ostreatus grown on neem and mixed bed with P. pulmonarius also cultivated 
on neem and mixed bed (21.33 days, 21.00 days, 21.67 days and 20.33 days respectively).  
The longest mycelia extension per day at 4WCI was recorded in P. pulmonarius grown 
on neem (0.63 cm). This was followed by the same variety cultivated on mango (0.60 cm). The 
least was observed in P. ostreatus grown on mixed bed which was comparable to the same 
variety cultivated on cassia (0.42 cm and 0.44 cm respectively). At 8WCI, P. ostreatus grown on 
mango (0.72 cm) produced the longest extension per day. This was followed by comparable 
results from P. pulmonarius cultivated on neem and P. ostreatus grown on neem (0.69 cm and 
0.68 cm respectively). The least results were observed in P. ostreatus grown on mixed bed and 
cassia with P. pulmonarius cultivated on cassia (0.55 cm, 0.56 cm and 0.56 cm respectively). At 
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12WCI, the longest mycelial extension per day was recorded in P. ostreatus cultivated on mango 
(0.76cm) but not significantly different from P. pulmonarius grown on neem (0.75 cm). This was 
followed by P. pulmonarius grown on mango (0.69 cm) which also was not significantly from 
the results of P. ostreatus cultivated on both neem and cassia (0.71 cm and 0.68 cm 
respectively). The least was however, observed in P. pulmonarius cultivated on cassia (0.59 cm). 
4.3 Effects of different substrate types on the yield of the sclerotia of Pleurotus tuber-
regium. 
 At the different stages of this study, all the sawdusts supported the production of sclerotia 
of Pleurotus tuber-regium (Plates 4.6 and 4.7). Generally, the highest sclerotia weights were 
recorded on the sawdusts with the longest weeks of composting interval (WCI) (Table 4.14). 
At 4WCI, the highest weight was recorded in Pleurotus tuber-regium sclerotia obtained 
from the sawdust of mango (23.24±1.81 g) followed by comparable results from cassia and neem 
sawdusts (17.30±1.59 g and 15.51±1.55 g respectively) while the least was on the mixed bed 
(9.00±0.60 g). At 8WCI, the highest weight was from P. tuber-regium sclerotia harvested on 
cassia sawdust but was not significantly different from the one grown on mango (35.34± 1.46 g 
and 32.42±1.48 g respectively).  
The least was recorded in mixed bed (16.94±1.40 g) which however was not significantly 
different from what was obtained on the sawdust of neem (18.98±2.50 g). Furthermore, mango 
sawdust produced the heaviest sclerotia at 12WCI (42.13±0.85 g) followed by the mixed bed 
while neem sawdust produced the least (37.02±1.71 g and 26.77±1.79 g respectively). 
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Table 4.13. Effects of substrates x varieties x weeks of composting interval (WCI) interaction on the growth parameters 
                 of Pleurotus ostreatus and P. pulmonarius. 
 Substrate   Full Mycelia Colonization (days)    Primordial Initiation (days)      Extension per Day (cm) 
 WCI (Weeks)    4     8    12     4     8    12    4    8    12 
 Mango 23.67ab 22.00b 17.00d 28.00ab 26.00b 20.33c 0.55c 0.72a 0.76a 
P ost Cassia 24.67a 23.67a 20.33ab 29.33a 27.00ab 23.33b 0.44f 0.56e 0.68b 
 Neem 25.33a 21.67b 19.67b 30.33a 25.00b 21.33c 0.47e 0.68b 0.71b 
 Mixed Bed 23.67ab 24.00a 18.67c 23.33c 26.33b 21.00c 0.42f 0.55e 0.64c 
           
 Mango 24.33ab 23.00ab 20.33ab 28.33ab 27.33ab 23.33b 0.60b 0.64c 0.69b 
P pul Cassia 25.67a 23.67a 23.00a 30.00a 28.00a 26.00a 0.52d 0.56e 0.59d 
 Neem 26.00a 24.67a 21.67a 29.67a 29.33a 21.67c 0.63a 0.69b 0.75a 
 Mixed Bed 25.33a 23.67a 20.33ab 29.00a 27.67ab 20.33c 0.57c 0.61d 0.65c 
 
Means with the same letter along the column are not significantly different from one another at p ≤ 0.05. 
P ost: Pleurotus ostreatus 
P pul: Pleurotus pulmonarius 
WCI: Weeks of composting intervals 
 
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Plate 4.6: Sclerotia of  P. tuber-regium on the substrates 
 
 
 
 
 
 
 
 
 
 
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            Plate 4.7: Harvested sclerotia of P. tuber-regium 
A: At 4 weeks of composting interval  
B: At 8 weeks of composting interval  
C: At 12 weeks of composting interval  
 
 
 
 
 
 
 
 
 
 
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Table 4.14. The interaction effects of substrate types and weeks of composting intervals 
                   (WCI)   on the yield (g) of the sclerotia of Pleurotus tuber-regium 
Substrates                                          Weeks of Composting Intervals 
 4 8 12 
Mango 23.24a 32.42a 42.13a 
Cassia 17.30b 35.34a 32.83c 
Neem 15.51b 18.98b 26.77d 
Mixed Bed 9.00c 16.94b 37.02b 
 
Means with the same letter along each column are not significantly different from one another (p 
≤ 0.05). 
 
 
 
 
 
 
 
 
 
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CHAPTER FIVE 
          DISCUSSION 
The rapid growth and the ability to utilize various lignocellulosic substances make 
Pleurotus species cultivation possible in different parts of the world. Pleurotus species have been 
grown on different kinds of sawdust, straw and many other agricultural and industrial wastes 
(Hadder et al., 1993). Some of these otherwise valueless lignocellulosic wastes: cotton wastes, 
sawdust, cereal stover, corncob, wheat, paddy straw and sugarcane bagasse, have been used 
either mixed or singly as substrates for the cultivation of various species of edible mushrooms by 
various researchers (Fasidi and Kadiri 1993; Manzil et al., 1999; Ragunathan and Swaminathan 
2003). They can colonize and produce mushrooms on pretreated conifer (Pinus spp.) wood chips 
but they do not always readily colonize non-pretreated conifer wood, due to the presence of 
inhibitory components (Croan, 2004). Some strains can, however, be adapted for cultivation on 
conifer sawdust-based substrates (Ruan et al., 2006). Pleurotus spp. can also be cultivated on 
wood waste or unused wood residues associated with harvesting or thinning operations, which 
can enhance economic returns needed to support ecosystem management (Croan, 2000). They 
have extensive enzyme systems capable of utilizing complex organic compounds that occur as 
agricultural wastes and industrial by-products (Baysal et al., 2003). 
Different substrates for cultivation have significant effects on the mushroom yield. 
Different yield amounts were obtained from varied substrate media. This result is in consonance 
with previous research findings of other researchers who reported various values for the yield 
(Ohga, 2000; Ragunathan and Swaminathan, 2003; Yildiz, 2003). Using varied substrate media 
for the cultivation of mushroom causes different yield amount because of the biological and 
chemical differences between the substrates medium and genotype of the cultured mushroom as 
previously observed (Imbernon, 1990; Olivier 1990).  
Number of fruits, fruit weight, the width of pileus, biological and production efficiencies 
increased as weeks of composting interval (WCI) increases in all the treatments. The release of 
nutrients was constant in mango and mixed bed substrates, increased in neem but decreased in 
cassia. This could probably be due to the fact that the longer the substrates are allowed to 
decompose, the more the nutrients that will be released and this might have enhanced the 
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increase of these parameters. However, the release of inhibitory substances might be atributable 
to the decrease in the yield parameters of neem. 
 The highest number of fruits and fruit weights, largest width of pileus, longest length of 
stipe, greatest biological efficiency (BE) and production efficiency (PE), shortest number of days 
for full mycelia colonization and primordial initiation with the fastest mycelia extension per day 
were recorded at 12WCI in the varieties. This could be attributed to prolonged decomposition of 
the components of the various sawdust used thus, making them available for the mushroom 
mycelia. The average fruit weight was however highest at 4WCI. It was observed that few 
numbers of fruits were produced at 4WCI resulting in the very high average fruit weight as 
against the emergence of more fruits as WCI increases. Various mushroom fruit weight values 
were recorded for the sawdust from the different wood types. This was similar to the report by 
Abott et al. (2009) where it was observed that that the yield of Lentinus squarrosulus was 
influenced by the type of sawdust used for cultivation. This means that saw dust type influences 
the yield of mushrooms. 
 Sawdust (wood) has been the traditional substrate for the growth of mushroom (Onuoha, 
2007). Sawdust has been consistently reported to be the best substrate supporting mycelia growth 
and fruitification (Kadiri and Fasidi, 1990). It was observed in this study that mango tree sawdust 
produced the greatest number of fruits though not significantly different from that of mixed bed 
and neem while the least was obtained from cassia. It was also noted that the fruit weight 
recorded for mango significantly followed that of mixed bed and cassia however, it produced a 
significant width of pileus, length of stipe, production efficiency with the fastest mycelia 
extension per day.  
Very high mycelium density was observed in mango sawdust, being a soft wood, 
resulting in very high yield. This could be due to the physical nature, high level of aeration and 
porosity within the substrate. Findings of Thomas et al. (1998) revealed that the yield of 
mushroom is directly related to the spread of mycelium into the substrate. Furthermore, values of 
BE observed varied as a result of the biological structure of the raw materials or substrates used 
in this study as previously reported  (Akyüz and Yildiz, 2007, 2008). The variations may also 
stem from the difference in the nutrient content of the materials. 
 Of the three mushroom varieties under investigation, greater effects were observed in P. 
ostreatus in terms of number of fruits, fruit weights, biological efficiency, production efficiency, 
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shorter days to full mycelia colonization and primordial initiation. This is attributable to the soft 
nature of mango tree. This agreed with the work of Hami (1990) that observed that P. ostreatus 
gave maximum biological efficiency on the sawdust of soft wood like mango. 
Generally, it was observed that all the parameters were significantly highest at 12WCI 
except mycelia extension, days to full mycelia colonization and primordial initiation. The 
increased mushroom yield must have been due to the prolonged substrate decomposition period, 
causing increased decomposition and making more nutrients available in the substrate for the 
mushroom growth. This was in agreement with the research findings of Rohrer and Heimlich 
(1989) that compared the decomposition rates of shredded newsprints, straw and saw dust. The 
authors observed that as the period of decomposition increases, the materials under investigation 
became more decomposed. After five weeks, the sawdust and shavings bedding material tended 
to blend with the dry surface of the soil, sawdust that was in a moist condition crumbled readily 
and nearly disintegrated when rubbed between the thumb and forefinger. The fine shavings and 
sawdust material decomposed most readily. As these materials were kept moist, they completely 
decomposed at the end of the 16 weeks test period. The authors observed that newsprint and 
straw were more closely paired in decomposition rates. The newsprint did decompose or at least 
disintegrate more quickly than straw when kept in similar moist conditions on the soil. The straw 
was most persistent in retaining its color and strength because of the long fibers. At the end of 
the 16 weeks study period, the straw was still yellowish in color and tore very easily. Most of the 
newsprint and straw were not decomposed. However, my investigation spanned only 12 weeks 
of composting interval. 
At 4WCI, the fastest mycelia extension was observed in neem. This could be as a result 
of minimal decomposition causing a sporadic spreading of mycelia because of the little available 
nutrients. The same reason is attributable to full mycelia colonization resulting in the shortest 
period for primordial initiation at 4WCI. 
 As observed, all the growth parameters revealed that P. ostreatus performed better than 
P. pulmonarius as WCI increases. This was indicated by the number of fruits, fruit weight, 
biological and production efficiencies and average extension per day, all at 12WCI. Also, as 
observed earlier, the yield of P. ostreatus was more than that of P. pulmonarius. This could stem 
from the resultant total yield harvested on mango sawdust, a soft wood. This was also reported 
by Hami (1990). 
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 Although neem sawdust produced the greatest number of fruits but it was not 
significantly different from what was obtained from mango. Of all the substrates under 
investigation in this study, mango sawdust produced the greatest yield with P. ostreatus 
performing better than P. pulmonarius. Furthermore, the biological and production efficiencies 
of mango were highest with P. ostreatus. This could have resulted from heavy mycelia 
ramification on mango sawdust because of its porosity and aeration as a soft wood. Mehravaran 
(1993) observed that active mycelia growth is directly proportional to maximum of respiration as 
oxygen (O2) is one of the most important environmental factors. It plays a major role in the 
metabolism and respiration of mushrooms giving rise to a better yield.  
However, both the mycelia extension per day and total mycelia extension of P. ostreatus 
were least in mixed bed and mango sawdusts respectively. Similar observation was made by 
Elhami and Ansari (2008) while conducting a research on the effects of substrates on spawn 
production with respect to mycelia growth of oyster mushroom species. They observed that 
mycelia growth was significantly affected by species (mushroom varieties). They stated that the 
best mycelia growth was obtained from P. florida followed by P. citrinopileatus while the least 
was from P. ostreatus. Nandi and Mukherjee (2004) also reported that P. florida was more 
effective than P. citrinopileatus in delignification while P. ostreatus also had the least growth 
rate. Furthermore, Smith and Margarel (1995) obtained similar report for Agaricus strain (W4II) 
while Jonathan and Fasidi (2003b) equally reported for Psathyrella atroumbonata.  
 It was generally observed that as WCI increases the yield also increases. At 12WCI, the 
number of fruits of P. ostreatus harvested was greater than that of P. pulmonarius. The fruit 
weights from both mango and cassia sawdusts were highest but not significantly different from 
each other. However, mango, as a substrate, performed best in terms of biological and production 
efficiencies and with the longest average extension per day. This could have been as a result of 
the temperature of the fruiting house, moisture level and compost preparation. The temperature 
of the fruiting house was kept low as the mushrooms were grown under controlled conditions. 
Wetting was applied when necessary to keep the relative humidity of the fruiting house very 
high. When the temperature and moisture level are at the best, maximum number of pinheads 
and mushrooms are formed. Compost preparation includes picking unwanted materials from the 
substrate (sawdusts), moistening with water and the duration of composting thereby releasing the 
nutrients for the maximum growth of the mushrooms. Rohrer and Heimlich (1989) stated that as 
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the period of decomposition increases, the materials (substrates) under investigation become 
more decomposed. 
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CHAPTER SIX 
CONCLUSIONS  
From the result, it may be concluded that mango sawdust is the most suitable for the 
production of the fruiting bodies P. ostreatus when compared with the other substrates. The use 
of this substrate gave the highest yields in terms of fruit number and weight, the width of pileus, 
length of stipe and mycelia extension. Thus, mango  is the best of all the substrates investigated 
and may be useful for large scale production of P. ostreatus. Furthermore, mango sawdust also 
gave the best in terms of the growth and yield of the sclerotia of P. tuber-regium. In comparison 
with other substrates under this investigation, cassia sawdust was most suitable and efficient for 
the production of P. pulmonarius. Full mycelia colonization took the longest number of days to 
maximize the cellulose and lignin contents availability resulting in the highest yields in terms of 
fruit weight, average fruit weight, production and biological efficiencies. 
The components of applied mushroom biology are closely associated with three aspects 
of wellbeing: food shortage, human health and environmental pollution. One of the most 
significant benefits of mushroom cultivation is their ability to create a pollution free and friendly 
environment. Mushroom is a short duration crop, its cultivation is land saving and can be 
welcomed by the poor farmers. The tropical trees usually constitute a nuisance in the 
environment, as they are allowed to rot naturally or set ablazed when cut down. This study has 
revealed that the sawdusts of mango, cassia and neem trees could be used to cultivate 
mushrooms. Thus, production of mushroom from tropical trees could compliment protein 
availability and a reduction in malnutrition. 
 
 
 
 
 
 
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UNIVERSITY OF IBADAN LIBRARY 
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