CHAPTER ONE

INTRODUCTION

1.1 BACKGROUND OF THE STUDY

Mushroom is the fruiting body of a particular group of macro fungi, which can be found growing
above or beneath the topsoil, seen macroscopically with the unaided eye and often collected for
food or medicine (Chang et.al 2008). Mushroom are highly valued for their distinctive taste and
nutritious nature. They are delicious food regarded as a good protein source and rich in vitamins,
minerals, and most essential amino acids (Gargano ML et.al 2017). In some continents, Europe
and Africa inclusive; mushrooms are usually collected from the wild and are used directly as
food or added to soups, stews, teas and can even be used as a replacement for meat. Mushrooms
are highly nutritious basidiomycetes that are artificially cultivated on lignocellulosic waste
materials. They are poor in fat and calories but known to be rich in minerals, protein and crude
fibre (Muthangya M et.al 2014). Mushrooms have a broad geographical distribution being the
earliest form of fungi that has been incorporated into cultural diets. In Nigeria, several people
living in the rural areas plus a few in the urban regions can identify edible mushrooms growing
in the wild, some of which have been exploited for income, food and medicine. Until recently,
many edible and medicinal mushrooms have been sourced from the wild forests since there are
very few commercial mushroom farms, especially in Nigeria. This could be partly due to less
craving for mushrooms due to mycophobic attitudes or preference for the naturally wild and
uncultivated species available for sale in some parts of the country. Mushrooms are yet to be
fully utilized due to low enlightenment, limited spawn availability (especially in Nigeria) and
poor preservation methods. However, regular mushroom consumption can supply essential
nutrients and prevent malnutrition. The mycorrhizal groups occurring in symbiotic association
with certain tree species such as Tuber melanosporum and Tricholoma matsutake. The second
group are commonly found in association with insects such as Termitomyces sp (Cheung 2008).
The third group exist as parasites deriving food substrates from living animals and plants, thus
causing a massive loss to these hosts. Examples of these include Coremiopleurotus, a tree
pathogen causing the white-rot of hardwood trees. The saprophytes obtain nutrients from dead
organic materials, including Pleurotus sp and Lentinus sp. There are well over 300 genera of
mushrooms and related fleshy basidiomycetes. Notably, in Nigeria, Osun State is endowed with
many Termitomyces sp. At the same time, Ore in Ondo state is home to Pleurotus tuberregium,
Agaricus sp., Marasmius sp., Termitomyces globulus, Coriolus versicolor and Fomes sp
(Gbolagade and Adeoye 2011). Some wild mushrooms are often available for sale in these
selected places seasonally. There are diverse mushroom species in the West, while Northern
Nigeria has very few if any at all. Tricholoma sp., Pleurotus sp., Lentinus sp., Psathyrella
atroumbonata, Schizophylum commune, Auricularia polytricha, Volvariella volvacea, Daldinia
concentrica, Daedalea elegans, Corilopsis occidentalis, Ganoderma sp. are some mushroom
species available in Nigeria (Gbolagade and Fasidi 2005), (Bamigboye et.al 2013).

1.2 STATEMENT PROBLEM

Producing mushroom spawn on cereals using spent substrate as a starter culture involves several
processes in achieving the targeted aim. However, challenges may arise from sterilization
processes, achieving consistent inoculation with spent substrate, and ensuring mycelium growth
for successful mushroom cultivation. These challenges contribute to the overall process of
spawning and production efficiency.

1.3 RESEARCH QUESTIONS

In the course of the research there are several questions the research will look into.

i. Examination of the effects of spent substrates on mushroom spawn production?

ii. Effects of spent substrates on mushroom spawn growth, yield and quality?
iii. What sterilization methods are most effective for cereals in mushroom spawn production?
iv. What are the factors that contributes to successful mycelium growth on cereals?

1.4 OBJECTIVES OF THE STUDY

The main objective of this study is to successfully cultivate mushroom spawn on cereals using
spent substrates as a starter culture. However, there are other objectives the research will provide
insight on.

i. To determine the impacts of spent substrate on the growth, yield and quality of
mushroom.
ii. To determine the most efficient and resource effective sterilization method for cereal in
mushroom spawn production.
 iii. To determine the factors that contributes to the growth of mushroom spawn on cereals
using spent substrate as a starter culture.

1.5 SIGNIFICANCE OF THE STUDY

The utilization of spent substrate, which is considered a free material, can significantly reduce
the cost of producing spawn compared to buying new substrate. By repurposing spent substrate
as a resource for new spawn production, this research contributes to a closed loop system that
minimizes waste and reduces the dependence on virgin materials. This study will play and
provide a key information on sustainable agriculture as well as white farming known as
mushroom farming.

1.6 JUSTIFICATION OF THE STUDY

This study will address the economic significance of optimizing mushroom spawn production on
cereals, offering potential cost savings for cultivators and contributing to the economic
sustainability of the mushroom. With a focus on sustainable practices, the study will promote the
effort of environmentally friendly agricultural practices. The study will add to the scientific
knowledge, by addressing the research gaps and contributing valuable information to the field of
mushroom cultivation, potentially paving way for further innovations and discoveries.
Mushroom cultivation provides a nutritious food source, and improvements in spawn production
contributes to enhancing food security by facilitating a more reliable supply of high-quality
mushrooms.
 CHAPTER TWO

LITERATURE REVIEW

2.1 HISTORY OF MUSHROOM CULTIVATION

Although, most fungi have a very short life span, they have existed for millions of years.
Imprints of mushroom lamellae have been found on wood dating back before the origin of man.
Hippocrates 470-400BC mentioned the medicinal value of mushrooms Galen AD130-200
advised that mushrooms should be picked only from field, grassland and meadows. During the
period of the Roman Empire their sale was regulated by law. For a long time before mushroom
production began in Europe and America, the Chinese had been growing mushrooms. The
species of Auriculara auricular was first cultivated in China in AD 600 while Flammulina
velutipes was grown around AD 800-900. Cultivation of other mushrooms such as Lentinus
edodes, Volvariella volvacea and Tremella fuciformis was first recorded in China in AD 1000,
1700 and 1800 respectively. Production of mushrooms in France by the inoculation (spawning)
of horse manure with spores from the wild mushrooms mycelia from the horse manure were used
to inoculate (spawn) new horse manure. Thereafter, some growers started to prepare composted
horse manure beds inside caves and disused mines and experimented with the use of
pregerminated spores or spawn to start rotten leaf litter was used as the casing layer. In 1779
Aberecromble described mushroom (Agaricus) production in England and Agaricus production
spread to other parts of Europe including Holland and Italy and later to Africa so that by the end
of the 19th century, a mushroom industry based on the cultivated of Agaricus bisporus started to
evolve in the western hemisphere, mainly in the United State and Western Europe. The rapid
development and growth of the mushroom industry from productive cave culture into one using
technical and controlled methods was stimulated in the 1960s as of now about 30 mushroom
species are cultivated and consumed as food with Agaricus bisporus being the mushroom
produced in the greatest quantity.

2.2 MUSHROOM CULTIVATION IN NIGERIA AND OTHER PARTS OF THE WORLD

Mushroom cultivation has great scope in Nigeria, China, and in some of other developing
countries because of the cheap and easily available raw materials needed for this activity,
coupled with faster means of communication and marketing (as a fresh commodity) and better
purchasing power of the people using China as an example, in 1978, the production of edible
mushrooms was only 60,000 tones but in 2006, China‘s mushroom production was over 14
million tones presently there are more than 30 million people directly or indirectly engaged in
mushroom production and businesses and China has become a leading mushroom producer and
consumer in the world whereas in 1997, Asia contributed 74.4% of the total world mushroom
tonnage, Europe 16.3% and North America 7.0%, both Africa and Latin America‘s shares were
less than 1% this is largely due to lack of know-how, lack of understanding that mushroom can
play vital roles towards enhancing human health when used as dietary food supplements, lack of
reliable sources of good quality mushroom spawn for supporting the efforts of local mushroom
growers, lack of venture capital to support mushroom farming entrepreneurs and absence of
systematic government support towards promoting mushroom farming as a valuable non-
traditional new food and cash crop (comparable to coffee, tea, cotton, tobacco, etc.). The
cultivation of mushroom in Nigeria has started gaining ground as people are now engaged in the
cultivation of mushroom and new methods of mushroom cultivation using locally available raw
materials currently on research by the Mycological Society of Nigeria (Mycoson) under the
Theme: ―Fungi and The Nigerian Economy” October, 2016 in Ebonyi State Federal University
Ndufu-Alike, Ikwo, in the following areas of research: i. Mushroom cultivation and Nigerian
economy ii. Mushroom cultivation/production and processing in Nigeria It is hoped that the
avocation of mushroom farming will become a very important cottage industry activity in the
integrated rural development programme which will lead to the economic betterment of not only
small farmers but also of landless laborers and other poor sections of the Nigerian communities.

2.3 MORPHOLOGY OF MUSHROOM

Mushrooms can be defined as “a macro-fungus with distinctive fruiting bodies, epigeous or

hypogeous, large enough to be seen with naked eyes and picked up by the hands”. The
mushroom fruiting body may be umbrella like or of various other shapes, size and colour.
Commonly it consists of a cap or pileus and a stalk or stipe but others have additional structures
like veil or annulus, a cup or volva. Cap or pileus is the expanded portion of the carpophore (fruit
body) which may be thick, fleshy, membranous or corky. On the underside of the pileus, gills are
situated. These gills bear spores on their surface and exhibit a change in colour corresponding to
that of the spores. The attachment of the gills to the stipe helps in the identification of the
mushroom.
2.4 MEDICINAL IMPORTANCE OF MUSHROOMS

The invention of the so called “wonder drug” penicillin was a landmark in the field of medicinal
uses of fungi. Since then, several fungi have been well recognized for their antifungal,
antibacterial, antiviral, antitumor and many others such properties of pharmacological values. In
the recent past a variety of medicinal preparations in form of tablets, capsules and extracts from
mushrooms have been produced and marketed.
Mushrooms are perhaps the only fungi deliberately and knowingly consumed by human beings
and they complement and supplement the human diet with various ingredients not encountered in
or deficient in food items of plant and animal origin. Besides, chemical composition makes them
suitable for specific group suffering with certain physiological disorders or ailments. Mushrooms
are regarded as an ultimate health food, low in calories due to presence of good amount of
quality protein, iron, zinc, vitamins, minerals and dietary fibres which protects from digestive
ailments and strengthening of the human immune system. Recent investigations have proved the
empirical observations of the oriental herbalists that certain mushroom possesses very useful
medicinal attributes. In the 1991, the value of world medicinal crops was estimated at 8.5 billion
dollars and in the same year 1.2 billion dollars are estimated to have been generated from
medicinal products from mushrooms. This was based on the sale value of products from
Coriolus, Ganoderma, Lentinula, Schizophyllum and other mushrooms. Although the biggest use
of mushroom has traditionally been for reasons of their gastronomic and nutritional appeal.
There has always been interest in certain mushroom for their medicinal attributes. Production of
medicinal mushroom is now a days increasing over world-wide. In the present era a variety of
proprietary product based on mushroom nutraceuticals and pharmaceutical have already been
produced and marketed. Various mushrooms and their metabolic extract have been reported to
protect against cancer, tumor and pathogenic microorganisms. It is suggested that regular
consumption of different mushroom varieties not only protects humans from heart trouble but
also had medicinal potential for certain ailments.

2.5 TYPES OF MUSHROOMS

They are two types of mushrooms edible mushrooms and poisonous. Edible mushroom includes
many fungal species that are either harvested wild or cultivated. Easily cultivated mushroom and
common wild mushroom are often available in market and those that are more difficult to obtain
may be collected on a smaller scale by private gatherers. Some preparation may render certain
poisonous mushroom fit for consumption. Before assuming that any wild mushroom is edible, it
should be identified. Proper identification of a species is the only safe way to ensure edibility.
Some mushroom that are edible for most people can cause allergic reaction in some individuals
and old or improperly stored specimens can cause food poisoning.

2.6 REQUIREMENTS FOR MUSHROOM CULTIVATION

2.6.1 Spore (Spawn)

What spawn is to mushroom is like seed is to crop. Unlike spore, spawn is already at its mycelial
stage growing on its own substrate such as sorghum, barley or sawdust. The life cycle of
mushroom starts from spores, but growers inoculate mycelialorigin spawn rather than spore
origin spawn because of possible variations and mutations. The quality of spawn is one of the
most decisive factors for successful crop. Therefore, growers need to use qualified spawn for
commercial production. Spawn should maintain the strain characteristics and is propagated by
subcultures. New strains are developed with genetic methods such as variation and mating. The
various types of mushroom spawn include grain, sawdust, plug and liquid (James 2009).

2.6.2 Substrate

Mushrooms can be classified into 3 categories by their tropic pattern; saprophytes, parasites or
mycorrhizae. The most commonly grown mushrooms are saprophytes, decomposers in an
ecosystem growing on organic matters like wood, leaves and straw in nature. Raw materials can
be used as substrate for primary decomposers such as oyster mushroom and enokitake. On the
other hand, secondary decomposers like button mushroom or straw mushroom require substrate
degraded by bacteria or other fungi. Mushroom requires carbon, nitrogen and inorganic
compounds as its nutritional sources and the main nutrients are carbon sources such as cellulose,
hemicellulose and lignin. Thus, most organic matters containing cellulose, hemicellulose or
lignin can be used as mushroom substrate. Examples are cotton, cottonseed hull, corncob,
sugarcane waste, sawdust, and so on. However, demanded amount of each nutritional sources
differs according to mushroom species. For example, button mushroom (Agaricusbisporus)
requires relatively high nitrogen source, so the optimal C/N ratio of button mushroom compost
is17. On the other hand, oyster mushroom and shiitake require less nitrogenand more carbon
source (James 2009). Mushroom mycelia secrete digestive enzymes into the substrate and absorb
the dissolved nutrients. Cellulose, the main nutritional source of mushroom is one of the most
abundant organic matters on earth, but its digestive enzyme, cellulase is owned by several
microorganisms including fungi. Here comes the reason mushroom is considered an important
food source. Mushroom is the only one by which cellulose is dissolved and absorbed and
transformed into food for mankind. Mushroom is also influenced by acidity of substrate. The
optimal pH value of substrate ranges from 6 to 8, varying with mushroom species (James 2009).

2.6.3 Environment

The last important factor for mushroom growing is providing an appropriate environment both
for vegetative and reproductive growth. Not being protected by a skin layer, fungi are easily
affected by their growing conditions. So, it can be said that the success or failure of mushroom
cultivation depends on the control of growing conditions. Environmental factors affecting
mushroom cultivation include temperature, humidity, light and ventilation. Optimal levels of
them at vegetative stage differ from those at reproductive stage. Mushroom mycelia can survive
between 5 and 40°Cdepending on the species. Mushroom mycelia grow well with the
temperature range between 20 and 30°C. Pins form at 10-20°C, lower than that of mycelial
growth by 10 °C. Over 80% of the fluid body is water. Substrate moisture content should be 60-
75% (James 2009). During fruiting, different relative humidity levels, ranging from 80-95%, are
needed at the early, mid and later stage. Though mycelia can grow without light, some species
require light for fruit body formation. Being aerobic fungi, mushrooms need fresh air during
growing, but ventilation is more required for reproductive stage. Among the three factors, the
most important is environmental control. By maintaining optimal conditions at each growing
stage and for each species, growers can produce the desired yield of quality mushrooms, (James,
2009).

2.7 ENVIRONMENTAL IMPACT OF MUSHROOM CULTIVATION

i. Reducing Environmental Pollution by Bioconversion: Organic solid wastes are a kind

of biomass, which are generated annually through the activities of the agricultural, forest
 and food processing industries. They consist mainly of three components: cellulose,
hemicellulose and lignin. The general term for these organic wastes is lignocellulose. It is
common knowledge that lignocellulosic wastes are available in abundance both in the
rural and urban areas. They have insignificant or less commercial value and certainly no
food value, at least in their original form. When carelessly disposed of in the surrounding
environment by dumping or burning, these wastes are bound to lead to environmental
pollution and consequently health hazards. It should be recognised that the wastes are
resources out of place and their proper management and utilization would lead to further
economic growth as well.
ii. Recycling of Organic Wastes into Mushrooms, Biofertilizer and Biogas: The ultimate
aim in the applied aspects of any scientific endeavor is to integrate wherever possible the
various disciplines of science as well as the technological processes in order that
maximum benefits accrue from such efforts. Combined production of mushrooms, biogas
and biofertilizer from the rural and urban organic wastes should be one of the aims of
such integrated schemes that can eventually be put into profitable operation. Though the
conventional and established approaches towards the production of food, fertilizer and
fuel exist, the explosive growth of the population vis-à-vis the rapid depletion of
conventional fuel resources leads mankind to look for alternative sources for food,
fertilizer and fuel.
iii. Restoration of Damaged Environment by Mushroom Mycelia: Mushroom cultivation
technology is friendly to the environment. Mushroom mycelia can produce a group of
complex extracellular enzymes which can degrade and utilize the lignocellulosic wastes
in order to reduce pollution. It has been revealed recently that mushroom mycelia can
play a significant role in the restoration of damaged environments. Saprotrophic,
endophytic, mycorrhizal, or even parasitic fungi/mushrooms can be used in
mycorestoration, which can be performed in four different ways: mycofiltration (using
mycelia to filter water), mycoforestry (using mycelia to restore forests), mycoremediation
(using mycelia to eliminate toxic waste, and mycopesticides (using mycelia to control
insect pests). These methods represent the potential to create the clean ecosystem, where
no damage will be left after fungal implementation.

2.8 SPENT MUSHROOM SUBSTRATE

SMS (spent mushroom substrate) is the material which is left after growing mushrooms.
Mushrooms are ready for harvesting within 2-3 weeks and the substrate that is leftover is called
SMS. The characteristics of SMS differ from mushroom to mushroom and hence its composition
varies respectively. SMS can be reused again for cultivation of mushrooms but it is
recommended to change the substrate and grow a new crop. Before using SMS, the process of
pasteurization is done so that any type of pest or any type of pathogen that causes diseases in
mushrooms are killed and hence this can be used for different purposes. In some cases, pesticides
are also used in the process of cultivation but these chemicals decompose fast and hence the
chemical residue is less in these substrates and therefore it does not affect to that extent
(https://extension.psu.edu/). The amount of SMS produced is 3-5 times more than the mushroom
produced and it varies; for instance, in button mushroom it is 5 times, oyster 3 times and 3-8
times more in paddy straw mushroom. India produces 5, 00, 000MT of spent mushroom
substrate. Spent mushroom substrate has its nutritional value 1.51% of N, 3.77% of P and 0.61%
of K. Also, the hydraulic conductivity of SMS was 6.22m/h. The water holding capacity is
95.03%. The pH of SMS differs and it's almost between 7.28-7.75 and chloride plays an
important role in conductivity (Kumbhar et al., 2014).

2.8.1 Uses of spent mushroom substrate

Once the mushroom is harvested the leftover residue which is known as spent mushroom
substrate can be reused for various purposes. Some of the uses of SMS which is performed and
practiced by various research institutions are mentioned below:

a. Bioremediation: It is the process of removal of contaminants with the help of living

organisms like fungi, bacteria etc. SMS contain different substances such as lignin
peroxide, manganese peroxide and other compounds which are responsible for
degradation of compounds like hydrocarbons, also SMS has the ability to breakdown
xenobiotic compounds and also the adsorption capacity of SMS where it adsorbs organic
and inorganic contaminants (Mohseni and Allen, 1999; Singh et al., 2011; Bobadilla et
al., 2019).
b. As a mulch: SMS can be used as mulch which not only holds the water but protects the
seed from birds eating away. But in this case, it's not suitable for vegetables which are
 sensitive to salt content in the soil. Best work as a mulch during spring and summer
(Sagar et al., 2009; Zhu et al., 2013).
c. Crop production: Cultivation of mushrooms generally is done on organic substrates and
therefore it can be used for the production of crops but again the problem of high salt
concentration lies and therefore (Chong and Rinker, 1994) suggested trickle irrigation for
leaching out excess salt. SMS can be used for the cultivation of greenhouse plants, field
crops and also as “Manure”. In this case it is transformed to a liquid fertilizer and then
used on soil (Sagar et al., 2009; Becher et al., 2021).
d. Greenhouse crops: SMS can be used for different greenhouse crops as a growing media
for different seedlings. It can be used as an alternative for peat which is used as potting
mixtures in greenhouses. Also, it can be used for different flowers like marigold, easter
lilies, petunia etc. (Young et al. 2010; Zhang et al. 2012).
e. Field crops: Different studies show that the blending of SMS in the soil not only
increases the yield but also shows an increase in the nitrogen fixing bacteria. Also, many
studies show that for oilseeds crops the protein content of crops can be increased due to
SMS incorporation. Also, the quality of ear was improved in a research done in National
Research Center for Mushroom, Solan, HP (Sagar et al., 2007; Danny et al., 2017).
f. Nursery crops: SMS can be used for the production of different nursery plants like
shrubs and also for the germination of grass seeds and all these results in a good yield.
The best mushroom for this purpose is Agaricus and pleurotus spp. (Danny et al., 2017).
g. Food for animals and fish: Mushroom cultivation contains straw and grains that are
usually present as a feed for livestock and therefore recycling SMS as feedstock for
livestock has proved successful during different research works. These contain various
bioactive enzymes which act as antioxidants and anti-inflammatory agents which increase
the nutritional quantity of feedstock. Also, it was studied that SMS has a high
preservative quality of around 8 weeks when inoculated with required bacteria for the
feed and it could be directly fed to the livestock. These microbes are termed as direct-fed
microbes (DFM) (Kim et al., 2007 & 2008) [16] and this has resulted in an increase of
weight and feeding efficiency of livestock (Deshmukh, 2019; Fazaeli et al., 2014).
h. Reducing the effect of pesticides: Pesticides, when directly used on crops, results in loss
due to various environmental factors like volatilization, leaching, run-off due to heavy
 rain and this results in soil water and air pollution and thus this can be minimized with
the use of SMS as it has high adsorption capacity (Gavrilescu, 2017).
i. Reuse in the cultivation of mushrooms: Spent mushroom substrate can be reused again
with proper management and many researches have given good results. In one of the
research SMS is used as a casing material (casing is a process done after spawn run and
in this process a blanket of soil is covered on the cultivating substrate). This process is
important for the proper growth of mushrooms. In casing, mostly peat is being used, but
peat being an expensive as well as its low availability makes it difficult for the growers to
get a good casing material and this led to the reuse of SMS as casing material. But for
this SMS needs to be recomposed for a good period of time and this would result in the
lowering of EC and breakdown of vegetative matter into humic substances. Hence this
reuse of SMS was done for the cultivation of milky white mushroom (Farsi et al., 2011;
Rahman et al., 2017).

2.9 IMPORTANCE OF THE MUSHROOM SPENT SUBSTRATES AND ECONOMIC

BENEFITS OF MUSHROOMS

The environment contains large volumes of unused lignocellulosic by-products from

anthropogenic activities or by natural occurrence. In most case these lignocellulosic byproducts
are deposed off by burning or left to rot in the environment (Angadam et al., 2021). However,
they can be utilized for mushroom cultivation which can be of great value. Agricultural waste
constitutes valuable resources that can be used to contribute to the economy in various ways. For
example, substrates can be reused to produce mushrooms or used as high-quality waste to make
compost that can be used in gardens to grow vegetables like spinach, cabbage or lettuce (Ringer,
2017). The spent mushroom substrate can be used as an alternative for energy production, to
produce biofuels or can be used as firewood (Carrasco et al., 2018). In addition, used substrate
can be used as animal feed to boost their health status as spend mushroom substrate is rich in
nutrients. Spent substrate can also be used for bioremediation to remove unwanted
contaminations in the air, soil and water (Ringer, 2017). Mushroom cultivation can easily be
carried out by unskilled farmers and waste are used and turned into products that are edible and
of high market value. Mushroom cultivation can contribute to food security and also can be an
alternative to employment especially disadvantaged groups specifically women and disabled
people (Tesfaw et al., 2015). Mushroom cultivation can be regarded as an important commercial
asset because it has potential to generate income. Varieties such as Agaricus bisporus, Lentinus
edodes and Pleurotus species are the most commercially and successfully cultivated mushroom
(Owaid et al., 2015).

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