YIELD AND SOME MORPHOLOGICAL CHARACTERS OF PLEUROTUS PULMONARIUS (FRIES) QUEL FRUIT-BODIES CULTIVATED ON HCL-OPTIMIZED OIL PALM BUNCH SUBSTRATE

ABSTRACT

The study was conducted to optimize the pH of oil palm bunch (OPB) as a substrate for the cultivation of Pleurotus pulmonarius fruit bodies. The natural pH of OPB was found to be 9.5. Hydrochloric Acid (HCL) made into solutions of 0.1%, 0.2%, 0.3% and 0.4% was used to alter the OPB pH at 8.9, 8.2, 7.9, 6.2 and 9.1 (control) after soaking for 48hrs. The result shows that 0.4% HCL treated OPB gave the highest yield (g) and Biological Efficiency (B.E%) of 900g and 90% respectively while control yielded the lowest (493g) and B.E of 49.3%. Result from the morphological characters of individual mushroom fruit body shows that Cap Size (C. S cm) and Weight (wt. g) was also the highest (3.83cm and 3.5g) in 0.4% HCL treated OPB while Stipe Length (S.L cm) was also high (2.77cm) in 0.3%. Morphological characters of P. pulmonarius fruit bodies showed no significant different at p > 0.05. There is also a high level of relationship between Stipe Length (S.L) and Cap Size (C.S) of individual fruit bodies and also C.S and Weight (Wt.) but no significant relationship between S.L and Wt. In conclusion HCL was found as a suitable acid buffer for the optimization of OPB substrate in the cultivation of P. pulmonarius fruit bodies and should be adopted for the commercial production of the mushroom as soon as the mushroom is certified safe for consumption.

TABLE OF CONTENTS

Title Page

Certification

Declaration

Dedication

Acknowledgement

Table of contents

List of Tables

List of Plates

Abstract

CHAPTER ONE

Introduction

1.1   World Mushroom Cultivation

1.2   World Mushroom Market

1.3   Taxonomy

1.4 Economic Important of Oyster Mushrooms

1.5 Nutritional and Food Values

1.6 Medicine/Tonic

1.7 Income and Job Creation

1.8 Justification

1.9 Objectives of study

CHAPTER TWO

Literature Review

2.1.   Effect of pH on mushroom fruit body formation

2.2.   Mushroom Cultivation

CHAPTER THREE

MATERIALS AND METHODS

3.1 Source of materials

3.2 Location of Study

3.3 Spawn production / multiplication

3.4 Determination of substrate pH

3.5 Substrate preparation

3.6 Substrate inoculation     

3.7 Measurement of morphological parameter

3.7.1Stipe Sizes of Fruit   Bodies

3.7.2 Diameter of Pileus

3.7.3. Fruit Body Weight

3.8   Yield and Biological Efficiency

CHAPTER FOUR

4.1   RESULTS

CHAPTER FIVE

 Discussion and Conclusion

5.1 Discussion

5.2 CONCLUSION

5.3 Recommendation

      Reference

Appendixes

APENDIX 1

APENDIX 2

ANOVA TABLE FOR S.L, C.S AND WTOF P. PULMONARIUS CULTIVATED ON HCL BUFFERED OPB.

LIST OF TABLES

Table1: Effect of percentage HCL conc. On substrate pH and spawn run duration of P. pulmonarius across the substrates

Table2: Effect of percentage HCL solution conc. on the yield and biological efficiency of P. Pulmonarius sporophores as affected by different HCL treated substrates

Table3:  Effect of percentage HCL solution treated substrate on S.L, C.S, and Wt. of fruit bodies

Table4:  Correlation effect HCL treated substrates on S.L, C.S and Wt. of P. pulmonarius fruit-bodies

LIST OF PLATE

            Plate 1: Fully Colonized Spawn of P. pulmonarius Ready for Inoculation.

           Plate 2: Oil Palm Bunch (OPB) Substrate.

          Plate 3: Oil Palm Bunch (OPB) Substrate Being Soaked In HCL Solution.

          Plate 4: P. Pulmonarius Fruit Bodies Growing in HCL Treated OPB Substrate.

CHAPTER ONE

INTRODUCTION

Mushrooms belong to the class Basidiomycota and order Agaricales. They do not possess chlorophyll like green plants for manufacturing their food but for their growth and development they require preformed food like smaller broken down molecules of cellulose and starch (Banjo, 1998). Chang (1991) defined mushroom as “a macro-fungus with a distinctive fruiting body which can either be epigeous (growing on or close to the ground) or hypogenous (growing under the ground)”. The macro-fungi have fruiting bodies large enough to be seen with the naked eye and to be picked up by hand. Ideally, the word mushroom refers only to the fruit body. Unlike green plants, mushrooms are heterotrophs, and without chlorophyll, they cannot generate nutrients by photosynthesis, but instead take nutrients from outer sources. Most mushroom species are either under the Basidiomycota or Ascomycota; the two phyla are under the kingdom Fungi (Cho, 2004).

In its simplest form, the life cycle of a mushroom may be traced from - a spore which under favorable conditions germinates to form a mass of branched hyphae of mycelia with colonies in a substrate. This represents the vegetative stage of its growth. When a given substrate is fully colonized, the vegetative growth ceases. Typically some of hyphae form primordial or fundament which is the beginning of the productive stage (Bahl, 1985). This develops further to form the stipes (stalk), the piles (cap) of the fruit body, which when mature exposes the gill, tissue or generative tissue on the underside, from which spores are liberated, so that the life cycle is perpetuated. Many fungi that form mushroom exist in mycorrhizal with trees, and this is one of the reasons why the forest is often the target for mushroom hunters. Many have learnt through the ages, by trials and error, to identify the edible and inedible mushrooms. In many cases some inedible ones resemble the edible types and are eaten without adverse effect (Gyar, 2007). However, there have been occasional accidents of consuming poisonous species leading to death or serious illness. Mushrooms have now been recognized universally as food and are grown on commercial scale in many part of the world including Nigeria. Gyar (2007­) observed that this fungus is common in Nigeria and often found growing around the African breadfruit (Treculia africana). In Nigerian, the most prized edible species are Pleurotus, Termifomyces, Tricholomaand Volvariella.

 Human population expands by 2.1% representing a rise of about 75 million people per year, thus food production has to keep pace with population increase (Sharma, 2003). Mushroom along with yeast are referred to as alternative source of food. (Chang, 1981; Anyankorah, 2002; James, 1995).According to James and Klegnhans (1995), edible mushroom (dry) contain about 19-40% protein; that is, its protein content is twice that of vegetable and four times that of oranges, and they are rich with vitamins, and minerals, less percent of unsaturated fatty acid and carbohydrate which makes it so ideal for diabetic and the obesity patient. Most mushroom has exceptional medicinal potentials and properties; curative and prophylactic especially in diseases such as high blood pressure, asthma, respiratory tracts infection, anaemia, hepatitis, cancer, tumour, etc. (Ogundana et al., 1982; Pelczar et al.,1986; Park et al., 2001) Mushrooms cultivation also serves as the most efficient and economically -viable biotechnology for the conversion of long-cellulose waste materials into high-quality protein food and this will naturally open up new job opportunities especially in rural areas and may be prepackage by food industry and exported to other countries as food conditions and for revenue generation. Pleurotus is the scientific name for Oyster mushroom. In many parts of India; it is known as Dhin ( Gyar, 2007; Ogundana et al., 1982).It belongs to the family Tricholomataceae which includes many species such as P. flobellotus P. sojar - caju, P. eryngii, P. osfreafies, P. floride and P. sapidus.

1.1 WORLD MUSHROOM CULTIVATION

Pleurotus species, commonly known as oyster mushrooms, are edible fungi cultivated worldwide especially in south East Asia, India, Europe and Africa. China produces 64% of all edible mushrooms in the world and 85% of all oyster mushrooms all over the world (Pleurotus spp.) is also produced in China (Chang, 1997). Oyster mushrooms is the third largest (Obodai et al., 2003).Commercially produced mushroom in the world; however, Sánchez (2010) reported that P. ostreatus is the second largest next to Agaricus bisporus in the world market. Mushroom cultivation is the fifth largest agricultural sector in China with 24 billion USD value and 10% growth rate every year for the last 30 years (Zhang et al., 2004). Oysters are naturally found on rotten wood material. The growing and consumption interest of oyster mushroom is increasing largely due to its taste, medicinal and nutritional properties (Garcha et al., 1993). Large volumes of unused lignocellulosic by-products are available in tropical and sub-tropical areas. These by-products are left to rot in the field or are disposed-off through burning. Utilizing these by-products for mushroom cultivation using locally available technologies may be one of the solutions to transforming these inedible wastes into accepted edible biomass of high market value. P. pulmonarius is commercially cultivated in New Zealand and sometimes sold as oyster mushrooms (Baggio et al., 2010). Presently, sawdust is the major substrate used in commercial cultivation of P. pulmonarius in Nigeria. Due to technology advancement, the so called waste (sawdust) is now been used greatly for the production of briquette, shelf, board, office table and furniture generally. The on-going publicity of mushroom as a high source of protein with low cholesterol content which over ride meats and other fatty foods, may soon diminish due to the fore-scarcity of the sawdust (Quimio et al., 1990).

1.2 WORLD MUSHROOM MARKET

The world mushroom market for the mushroom industry in 2001 was valued at over US $40 billion. The mushroom industry can be divided into three main categories; edible mushrooms valued about US $30 billion; medicinal mushrooms products were worth about US $9-10 billion and wild mushrooms, US $4-5 billion (Chang, 2013). These mushrooms are traded mostly in processed form but lately, fresh mushrooms are being preferred over preserved ones in Europe and American countries (NRCM, 2004). Major exporting countries of fresh mushrooms are Netherlands, Poland, Ireland and Belgium (Chandy, 2010). China is the largest exporter of preserved mushrooms with the market share of 41.82% (more than 1.5 million metric tons in 2009) and 4.9 million metric tons in 2011, they are here by maintaining her position on the lead table (FAO, 2011). Netherlands (25.11%), Spain (7.37%) and India (4.44%) are the major producing countries (NRCM, 2004).

The major importing countries of prepared and preserved mushrooms are Germany, USA and France, while of fresh mushrooms are UK, Germany, USA and France (NRCM, 2004; Chandy, 2010).

1.3 TAXONOMY

Pleurotus  pulmonarius (Jacq.) P. kummer, is an edible white rot fungi (WRF) commonly known as the Indian Oyster, Phoenix Mushroom, or the Lung Oyster is classified as follows:

Kingdom         Fungi

Division          Basidiomycota

 Class  Agaricomycetes

Order   Agaricales

Family Pleurotaceae

Genus  Pleurotus

Species: P. pulmonarius. (Jacq) P. Kummer. Species of Pleurotus have been recorded and new species are discovered more or less frequently although some of these are considered identical to previously recognized species. The genus Pleurotus, which was first recommended as a tribe within genus Agaricus by Fries (1821). Three genera of this group, Pleurotus, Lentinus, and Panus, were possible to be separated according to their anatomic characters of the sterile tissues of the hymenophores as being homogeneous taxonomic groups. Hilber (1982) recommended that crossing of mono-spore cultures is a valuable basis for Pleurotus studies. Pleurotus pulmonarius (Jacq: Fr.) Kummer is the most cultivated species among the oyster mushroom and the type species of the genus Pleurotus. Recently, the majority of mycologists have followed the proposition made by Singer (1986) which divides the genus Pleurotus into six sections: Sect. Lepiotarii (Fr.) Pilat, Sect. Calyptrati Sing., Sect. Pleurotus Sing., Sect. Coremio Pleurotus (Hilber), Sect. Lentodiellum (Murr.) Sing. And Sect. P. tuberegium Sing. Pleurotus pulmonarius was placed in the Sect.

1.4 ECONOMIC IMPORTANCE OF OYSTER MUSHROOMS

The economic importance of oyster mushrooms can be described based on the following.

1.5 NUTRITIONAL AND FOOD VALUES

The desirability of a food product does not necessarily bear any correlation to its nutritional value, instead, its appearance, taste and aroma, sometimes can stimulate ones appetite. (Chang, 2013) in addition to nutritional value, mushrooms have some unique colour, taste, aroma and texture characteristics which attract their consumption by humans (Sabir et al., 2003).

Mushrooms are consumed for nutritional as well as their food values (Agraher-murugkar and Subbulakshmi, 2005).  Pleurotus spp. Are among the edible mushrooms consumed in the tropical states of West Africa and it is used as substitutes for meat and fish in some cases (lwalokun et al., 2007). Apart from being known for their appetizing flavor, they also offer themselves as potential protein, mineral and vitamin sources (Wahlid et al., 2006; Chang, 2013). Mushroom protein is intermediate between that of animals and vegetables, but superior to most other foods, including milk and contains all the nine essential amino acids required by man (Parkayastha and Nayak, 2002; Chang and Miles 2004; Kurtzman, 2009). Mushrooms contain appreciable quantities of crude fibres, although little information exists on the total dietary fibre (TDF) contents of mushrooms. Crude fibre content values reported by many authors suggest that mushrooms are potential sources of dietary fibre (Kurasawa (2004); Furlani, 2004). Okwulehie et al., (2008) reported high crude protein and carbohydrate contents in P. ostreatus var florida fruit bodies cultivated on different substrates and substrate supplementations.

According to Okhuoya and Okigbo (1991), Okwulehie and Odunze (2004a), mushrooms generally contain low-oil and fat, and because of the low content of oil and fat in mushrooms, they are recommended as good supplements for patients with cardiac problems.

The vitamins content of many mushrooms have been investigated and results of such investigation show that they are rich in vitamins including Thiamine, Riboflavin, Ascorbic acid, Ergosterine and niacin (Okwulehie and Odunze 2004a; Chang, 2013). Since vitamins are essential in the diet of man, and conventional sources of vitamins are scarce in recent times, it is pertinent therefore that attempts made to increase the list of the sources of cheap vitamins, is not misdirection (Aletor, 1995) and Fasidi (1996) have also reported that tropical mushrooms are rich in mineral nutrients and carbohydrates.

In Nigeria, P. tuber-regium is used as both food and medicine. The Sclerotium which is hard is peeled and ground for use in local cuisines (Okhuoya, et al., 2010). The mushroom itself is simply chopped and used in soups or may be dried for future use (Okhuoya and Okigbo, 1990). Wermer and Beelman (2002) reported that there has been a trend toward discovering ways of treating mushrooms so as to give them added value. For example, mushrooms enriched in selenium are now grown commercially.

1.6 MEDICINE/TONIC

For the past 20 years, interests in medicinal aspects of mushrooms have greatly been stimulated by the large number of scientific studies conducted on mushrooms (Tricita, 2004). Using modern approaches, scientists have isolated and identified specific components that can either destroy or at least debilitate three of mankind’s killer diseases: Cancer, heart disease and HIV/AIDS. (Tricita, 2004). The most recently introduced medicinal mushroom is Ganoderma spp. Its fruiting-body has traditionally been used for medicinal purposes and for thousands of years has been regarded by Chinese to be high quality herbal medicine (Oei, 2003; Tricita, 2004). Mushrooms have been used for ant-tumour, anti-cancer and many other therapeutic purposes. (Liu et al., 1995; Chang and Miles, 2004) Being rich in folic acid, mushrooms counteract pernicious anaemia (Oei, 2003). The polysaccharide-protein-complex (PSPC) found in mushrooms have proven to be anti-tumour, immune-modulatory, anti-malaria, anti-viral, anti-cancer etc. (Wang, et al., 2001; Liu et al., 1995). Cholesterol (a dreaded sterol for heart patients) is absent and can be converted to vitamin “D” by the human body (Chang et al., 2004). Nigerian native doctor’s use various combinations of herbs, mushrooms and other ingredients in their medicine. P. tuber-regium is used in some of these combinations that are intended to cure headache, stomach ailments, cough, cold and fever (Okhuoya et al., 2010). The considerable Pharmacological activities of mushrooms make them to be of interest in Pharmaceutical industries for the development of drugs (Okwulehie et al., 2008). Most bioactive compounds which play essential roles in human and animal physiology have been found in many mushrooms.

According to Okwulehie and Odunze (2004), Auricularia auricula, Pleurotus squarrosulus and Russula spp have been found to contain appreciable amounts of Alkaloids, Phenols, Saponins and flavonoids. In his investigation, Sofowara (2003) indicates that the plants widely used in traditional medicine contain in one or more of its parts potent bioactive compounds which are precursors for useful drugs synthesis.

According to Okwulehie et al., (2009), Fasidi, and Olorunmaiye .1994)), P. pulmonarius grown on various substrates gave low Alkal`oids, flavonoids, phenols and tannins composition. They noted that the values were lower than lethal doses for human. 

Alkaloids have powerful effects in animal physiology and are of interest in Pharmaceutical industries for compounding drugs (Edeoga and Eriata, 2001). According to Rambelli and Menini (2000), Alkaloids are stimulants and act by prolonging the action of many hormones.

Flavonoids have been reported to be useful in the treatment of some physiological disorders and diseases. According to Hilang and Feraro (2002), flavonoids have been reported to have anti-oxidant properties and are used as anti-carcinogens and ageing substances. In a similar way, flavonoids are said to have anti-bacteria functions (Dokara, 2006).

Tannins also inhibit pathogenic fungi and also reduce the rate of grazing on plants by animals that feed on plants. They also affect many human physiological activities such as stimulation of phagocyte host-mediated anti-tumour activities and a wide range of anti-infective actions (Haslam, 2006).

Saponins which are characterized by the soap-like foaming they produce when shaken in aqueous solutions have been implicated in the prevention of parasitic fungal diseases (Okwulehie et al., 2007).

1.7 INCOME AND JOB CREATION.

Growing mushrooms is labour intensive and for countries where jobs are scarce, Oyster mushrooms cultivation can create jobs, both in semi-urban and rural areas. (Won-sik, 2004). Tricita (2004) reported that some technologies can use family labour, thus providing all members of the family with employment. He went further to say that the labour of out-of-school youths and even school children can also be utilized especially as the bagging of substrates and related operations can be easily done by children.

Mushroom growing is also recommended as a project in a co-operative, where division of labour is practiced, one group will do the substrate preparation and still another group maybe engaged in spawn production and still another group can take charge of growing condition management (Tricita, 2004).

1.8JUSTIFICATION

pH is an important factor for good production of oyster mushrooms. Most mushrooms grow and perform well at pH near to neutral or slightly acidic at 6.1 and 7.5 respectively (Khan et al., 2013).  Oyster mushroom growers prefer to use lime (Ca Co3) as an alkaline buffer to optimize pH of some acid substrates.

Oil palm bunch (OB) is one of the most abundant organic materials often seen as waste around homes and oil palm meals. Although, before now, people living in the rural areas have used it to produce soap, though of low market quality. However, researchers have continued to search for the most effective innovation that would harness this abundant material either as a waste management strategy or for job creation.

Many growers of oyster mushroom have experienced difficulties in growing Pleurotus ostreatus using oil palm bunch. Achufusi (2016) used oil palm bunch as a substrate for the cultivation of P. ostreatus. The substrate was contaminated by Corprinus cinerius (a contaminant mushroom) and observed no yield of P.ostreatus fruit bodies in the substrate and its supplementations. The substrate pH was later found high at 10.3 and was suggested as the reason for Corprinus contamination and no yield of P .ostreatus.

In this investigation, HCL solution was used as an acid buffer to optimize the pH of oil palm bunch for the cultivation of Pleurotus pulmonaria fruit bodies.

1.9 Objectives of study

The aims and objectives of this work are to:

ü  Determine the pH of oil palm bunch (OPB) as a possible substrate material for cultivation of P. pulmonarius.

ü  Determine the yield and Biological efficiency of P. pulmonarius cultivated on Hcl treated OPB.

ü  To ascertain the level of relationship between the stipe length, cap size, and weight of mushroom fruit bodies.

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