PHYTOCHEMICALS, MINERALS AND HEAVY METALS EVALUATION OF PLEUROTUS OSTREATUS (JACQ) P.KUM FRUIT-BODIES CULTIVATED ON THREE STRAW SUBSTRATES

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ABSTRACT

The work involved the investigation of the effect of different substrates on the phytochemical, heavy metals and minerals var florida. The highest gross of heavy metals fruit bodies was obtained from the substrate Andropogon gayanus (351.75±0.30), followed by Pennisetum straw (342.41±0.20), while the least was found in Pannicum maximum (338.05±0.01). Similarly, the fruit bodies of substrates Andropogon gayanus weighed more than those from other substrates for both fresh and dry weight. Based on the phytochemical composition, Tannins was found to be highest (0.73±0.02) in mushroom harvested from A.gayanus followed by Pennisetum straw (0.66±0.02) while fruit-bodies harvested from P. maximum gave the lowest (0.55±0.03) phytochemical composition. Based on the mineral composition, result showed that all the fruit bodies obtained from A. gayanus substrates contained appreciable amount of calcium (1.60±0.30), potassium (2.45±0.10), magnesium (31.25±0.04), and sodium (2.30±0.30). Similarly fruit body obtained from P. ostreatus had highest (74.37±0.30) concentration of Phosphorus while significant difference in the amount of Sulphur containing fruit bodies harvesting in all substrates at p<0.05.  However, the lowest (0.70+-0.10) Calcium composition was recorded in mushroom harvested from P. purpureum. Likewise P.maximum  which gave the lowest of Sodium (1.60±0.20) and free Nitrogen (1.80±0.20). The result of the investigations was discussed in line of the best substrates for maximum production of Pleurotus ostreatus fruit-bodies with better qualities. The investigation concluded that growing Pluerotus ostreatus on A. gayanus  straw produced the highest fruit-bodies among other substrates which will help in the wide production of mushroom for market value. In general, Pleurotus ostreatus contains nutritional properties Also, the result from the investigation revealed that A.gayanus, Pannicum maximum, and Pennisetum straw could be vital in production of P.ostreatus from fruit bodies with A. gayanus supporting the highest P.ostreatus fruit-body production.People should incorporate the habit of eating mushrooms because of their high nutritional values.




 

 

 

 

TABLE OF CONTENT

 

Title page                                                                                                                                i

Declaration                                                                                                                             ii

Certification                                                                                                                            iii

Dedication                                                                                                                              iv

Acknowledgement                                                                                                                  v

Table of content                                                                                                                      vi

List of Tables                                                                                                                          ix

Abstract                                                                                                                                  x

CHAPTER ONE                                                                                                                   1

INTRODUCTION                                                                                                                1

1.1 Background of Study                                                                                                        1

1.1.1 What are Mushrooms                                                                                                     1

1.1.2 The Genus Pleurotus                                                                                                     3

1.1.3 Importance of Mushrooms                                                                                             4

1.1.3.1 Mushroom as Food                                                                                                     4

1.1.3.2 Nutrients in Mushrooms                                                                                             5

1.1.3.3 Vitamins                                                                                                                     7

1.1.3.4 Minerals                                                                                                                      7

1.1.3.5 Mushroom are Sources of selenium                                                                           7

1.1.3.6 Ergothioneine                                                                                                             7

1.1.4 Therapeutic benefits of Mushroom                                                                               7

1.1.5 Justification                                                                                                                   8

1.1.6    Aims and Objectives                                                                                                  9

CHAPTER TWO

LITERATURE REVIEW                                                                                                    10

2.1       Mycochemical Contents of Mushrooms                                                                     11

CHAPTER THREE                                                                         

MATERIALS AND METHODS                                                                                         14

3.1       Source of Mother Culture (SPAWN)                                                                         14

3.2       Location of Study                                                                                                       14

3.3       Experimental Design                                                                                                  14

3.4       Spawn Multiplication                                                                                                 14

3.5       Substrate Preparation                                                                                                  15

3.6       Spawn Inoculation                                                                                                      15

3.7       Sample Preparation                                                                                                    16

3.8       Determination of Heavy Metals                                                                                 16

3.9       Determination Of Mineral Content Of The Mushroom                                             17

3.10     Determination Bioactive Compounds In The Mushroom Fruit Bodies             17

3.10.1 Determination of alkaloids                                                                                         17

3.10.2  Determination of flavonoids                                                                                       17

3.10.3  Determination of phenols content                                                                              18

3.10.4  Determination of Saponins                                                                                         19

3.10.5   Determination of tannins                                                                                           19

CHAPTER FOUR

RESULTS AND DISCUSSION                                                                                           20

4.1       Results                                                                                                                        20

4.2.      Discussion                                                                                                                   23

CHAPTER FIVE

CONCLUSION AND RECOMMENDATION                                                                  25

5.1 Conclusion                                                                                                                        25

5.2 Recommendation                                                                                                             25

      References                                                                                                                        27

      Appendix  I                                                                                                                       32

      Appendix II                                                                                                                       36

 

 

 




 

 LIST OF TABLES

 

Table 41: Effect of straw substrate on the heavy metals                                                        20

Table 4.2: Effect of straw substrate on the minerals                                                              21

 Table 4.3: Effect of straw substrate on the phytochemicals                                                 22

 

 

 

CHAPTER ONE

INTRODUCTION


1.1 Background of Study

1.1.1 What are Mushrooms?

Mushrooms are the fleshy, spore-bearing, reproductive structures of fungi belonging mainly to the division Basidiomycotina and some members of Ascomycotina. According to Okwuliehie and Ogoke (2013) the fruit-bodies of mushrooms appear spontaneously in the forest and farm- lands in great quantities after the rains. The word “mushroom” is most often applied to those fungi (Basidiomycota and Ascomycete) that have a stem (Stipe), a cap (Pileus), and gills (lamellae, sing. Lamella) on the underside of the cap. Although they are plants they do not produce chlorophyll like green plants. They consist of stipe (Stem) that arises from underground mycelium and a Pileus or cap, with lower surface for spore development (Chang and Miles, 2000). They therefore do not manufacture their own food, but rely on substances such as litter, sawdust, decaying tree logs, wooden plugs, straw, and wood chips, or soil for nourishment. They require preformed food like smaller broken down molecules of cellulose and starch (Banjo, 1998).Mushroom can be found growing anywhere and on any natural material. They also grow on grassy ground, rotten leaf litter, dung, and other favorable substrates. They are widely distributed and have a wide range of shape taste and colour. Usually, the fruiting–bodies of mushrooms are umbrella shaped structures which produce spore in large numbers. These spores are minutes or microscopic and are dispersed by wind; under favorable condition they germinate to form a mass of branched hyphae or mycelia with colonies in substrates. This represents the vegetative stage of its growth. When a given substrate is fully colonized, the vegetative growth ceases.

These fruit-bodies which are known today as mushrooms appear white and have a short life span. 

In Nigeria, rural dwellers consume mushrooms as delicacies in soups and as ingredients for seasoning or part of the local melon cake (usu) in Igbo nation. Edible mushrooms are found in the wild especially with the onset tropical rainfall. In nature, a number of species of mushrooms are found mainly during the rainy season, on almost all type of soil, on decaying organic matter. Mushrooms have profound biological, ecological, and economic impact in the Nigeria and the world in general. The wild mushrooms which are edible, collected from the forests, gardens and other suitable substrates and are sold in the market of remote areas and also in the supermarkets of site life areas.

Many people have learnt since ages, (by trials and error) to identify the edible and inedible mushrooms. In many cases poisonous mushrooms are look-alikes of the edible ones and are often eaten without adverse effects (Bao, 2004).

Edible mushroom is a microfungus with a distinctive fruiting-body, which can be hypogeous or epigeous, large enough to be seen with unaided eyes. These mushrooms are grouped under Basidiomycetes but these are still some species of Ascomycetes that belongs to mushroom. As a whole, mushrooms constitute at least 14,000 and as many as 22,000 known species. However, the number of mushroom species in earth is estimated to be 140,000 suggesting that only 10% are known (Lindequisite et al., 2005).

Edible mushrooms have long and rich history of use. More than 2,000years ago, Paleolithic people knew and used mushrooms as powerful medicines to flight illness. In the world of pre-world Shaman, spirits caused disease and medicinal compounds were administered to appease or threat them. Mushroom was used as same tool as a practical treatment for microbial infection.

The medicinal use of mushroom has a very long tradition in the Asian countries, where as their use in West hemisphere has been slightly increasing only since last decades (Lindequisite et al., 2005). In Asia, mushroom in traditional Chinese medicine have been summation of more than 3,000years of use for the prevention and treatment of diseases (Chang, 2006). Historically, medicinal mushroom species harvested from the forest where they grew on dead or living trees and forest litter.

Edible mushroom have been considered to have medicinal value and to be devoid of undesirable effect. They may be eaten as meat substitutes and used as flavorings in foods, soups, and sauces. They also contain large amount of vitamins and minerals. (Oie, 2003, Okwulehie et al.,2007). In developing countries like India with rich biodiversity mushrooms are the bone for progress in the field of food, medicine and unemployment because of several nutriceuticals and medicinal mushrooms that have been found to be useful towards human health development as food, medicines, minerals and drugs among others (Wani et al., 2010).


1.1.2 The Genus Pleurotus

Pleurotus is a genus of gilled mushrooms which includes one of the widely edible mushroom species. Pleurotus is also called oyster, mushrooms comprises some of the commonly cultivated mushrooms in the world (chang, 2004). Pleurotus means “side ear”. The cap maybe laterally attached (with no stem), if there is a stem, it is normally eccentric and gills decurrently along it. The spores are smooth and elongated where hyphae meet. They are joined by clamp connections.

Pleurotus is not considered to be a bracket fungus and most of the species are monomitic (with a soft consistency). However, it can sometimes be dimitic, meaning that it has additional skeletal hyphae which give it a tougher consistency like bracket fungi (Knuden, 2008). Pleurotus fungi are found both in tropical and temperate climate throughout the world (Chang, 2004).

Classification of the species within the genus Pleurotus is difficult due to high phenotypic variability across wide geographic ranges, geographic overlap of species and ongoing evolution and speciation.More recently, molecular phylogeny has been tied to determine genetic and evolution relationship between groups within the genus, delineating discrete clades (Gonzalez, 2000, Zervakis, 2004). Pleurotus species are morphologically varied and are described as having convex or flat, fan shaped, kidney-shaped, asymmetric, appolaritic or sometimes funnel-shaped pileus with in volute margin (Rembelli and Menini, 1983). The size of the pileus ranges from 4.15cm in diameter, whitish, grey- white, violet grey, brownish grey, pink or violet black. (Rambelli and Menini, 1983). The gills are dense, unequal decurrent and light in colour. The oyster mushroom is commercially cultivated around the world for food, oyster mushroom can also be used industrially for mycoremediation purposes (Chang, 2004).it is one of the more commonly sought wild mushroom, it can be cultivated on straw and other media


1.1.3 Importance of Mushrooms

1.1.3.1 Mushroom as Food

The food and agricultural organization recognized mushroom as food containing protein nutrition to the countries depending largely on cereals (FAO, 2011). Mushroom with their flavour, texture and high productivity per unit have been identifies as an excellent food source (Eswaran and Rambadran, 2000). 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 one’s appetite ( ).Mushroom showcase the qualities expressed by the above statement. Mushroom has major unique colour, taste aroma and texture characteristics which attract their consumption by humans (Sh-ting, 2004).

Mushroom have been considered as source of rich food because they contain protein, sugar, glycogen, lipids, vitamins, amino acids and crude fibres (Okwulehie and Odunze, 2004a, Okwulehie et al., 2007, Okwulehie et al., 2009, Okwulehie and Ogoke, 2013). The protein of mushrooms is twice that of Asparagus and potatoes, four times that of tomatoes and carrot and six times that of oranges (Adejumo and Awosanya, 2005, Jonathan et al., 2006, Okwulehie, et al., 2014a). Apart from being a relatively rich source of protein compared to animal’s protein, mushroom could also hold special attraction and maybe recommended or people with cholesterol- related ailment (Adejumo and Awosanya, 2005).


1.1.3.2 Nutrients in Mushrooms

Mushrooms has been in consumption in the past centuries, but the discovery of the full nutritional values and other benefits from mushrooms is recent.Mushrooms have been found to contain all the essential amino acids that form the proteins.                       

Falade et al., 2008 observed that some mushroom species in Nigeria contain 81.4% and 93.2% moisture and also have some plant nutrients like tannin and trypsin inhibitor. In addition, mushrooms have been reported to accumulate several trace elements at greatly exceeding contents than in other plant food (Kelac et al., 2004). The protein content of mushrooms is about twice that of vegetables and four times that of oranges and more over the essential amino acids are present in mushroom. Crude protein was found in Pleurotus Ostreatus 27.3% and minimum P. djmor 24. 83% and P. sajor- Cajus 25.25% (Jawad  et al., 2010). Sadiq et al., 2008 reported the protein content of wild 24.66% in   P. Ostreatus. Similarly Okwulehie (2004) gave the protein content of 4 Nigeria mushrooms Auricularia auricular, Pleurotus Squarrosulus, P. tuber-regium and Russula spp range from 15.0 - 24.96g/100g on dry matter basis.

Mushroom is very rich in vitamins, they provide several types of vitamins, particularly thiamine (Mohan and Singh, 2011), Ekpo and Aluko, (2002). Riboflavin, Niacin, Biotin, Ascorbic acid, vitamin D and vitamin K, (Okwulehie 2004a).

On dry weight basis, Pleurotus Species 30-144mg/100g ascorbic acid, 1.16-4.8mg/100g thiamine, 109mg/100g niacin, and 4.7mg/ 100g riboflavin. Volvariella volvacea contain 206.27mg/100g of ascorbic acid while Nigeria Auricularia auricular, P. squarrosulus, P. tuber-regium and Russula species contains 5.46-9.86mg/100mg, 0.49-0.67mg/100mg riboflavin and 0.08-0.17mg/100g thiamin (Okwulehie and Odunze, 2004a, Okwulehie et al., 2009). Randive (2012) reported that oyster mushrooms are  rich in vitamin C, B complex required by the  human body.

Compared to cereals, pulses, fruits and vegetables the protein content of mushroom is much lighter on dry weight basis. Usually protein content of cultivated mushroom range from 2-5.3%, on fresh weight basis and 20-35%.On dry quality and of high die stability (70-83% properties). Cereals are deficient in two amino acids Lysine and tryptophan. But mushroom core rich in these two amino acids apart from most of the essential amino acid like leucine, isoleucine, valine, threonine, tyrosine and phenylamine. The order two essential amino acids, methionine and cysteine are however less in mushroom but in cereals (Tasvina et al., 2013).

 

1.1.3.3 Vitamins

Mushrooms are rich source of vitamins B complex (Thiamine, Riboflavin and Niacin) and VitaminC (ascorbic and acid) they also contain folic acid and Vitamin B12, which are not found in green vegetables.


1.1.3.4 Minerals

The minerals like potassium, sodium and phosphorus are abundantly found in mushroom. Which make it ideally suitable for hypertensive patients (Tasvina et al., 2013).


1.1.3.5 Mushroom are Sources of selenium

It is a mineral that works as an antioxidant to protect body cells from damage that might lead to heart disease, some concerns and other diseases of aging. It has also been found to be important for the immune system and fertility in men (Fresh – mushroom, 2014).


1.1.3.6 Ergothioneine

It is a naturally occurring antioxidant that also may help protect the body’s cell.Mushroom provides 2.8-49mg of ergothionine per serving of white, portabella or criminal mushrooms (fresh- mushroom, 2014). It is a mineral that works as an antioxidant to protect body cells from damage that might lead to heart disease, some concerns and other diseases of aging.


1.1.4 Therapeutic benefits of Mushroom

  1. Mushroom are  referred to as “vegetarian meat” as it  is rich in protein 35%
  2. Mushroom are suitable for diabetic  patients low carbohydrate content  and low starch
  3. Mushrooms are suitable diet for obsess person as these are low in calories (32 kcal/100g) fresh mushroom.
  4. They contain ergosterol which converts into vitamin D in the human body. The deadly cholesterol is absent.
  5. It is  high fibre content
  6. Mushrooms are beneficial for acidity gastritis problem due to presence of more than 7% alkaline ash.

1.1.5 Justification

According to Okwulehie, and Odunze, (2014a), the importance if mushrooms cannot be over emphasized. They are consumed on daily basis and are regarded as healthy additives to diets because they contain relatively high concentration of protein, fiber minerals and vitamins. Availability of suitable substrate for commercial production of Pleurotus ostreatus  within Umudike in Abia state has remained a relative problem, especially in the  availability of substrate such as rice straw which is best suitable for the  culture of P. ostreatus with the confirmation of Andropogon a yet another suitable substrate as stated by Okwulehie and Okwujiako (2008). Andropogon is one of the major grass plants in Umudike, it is not eaten as food but most times are cleared and burnt to ash either during farming or clean up exercises. It is however justifiable to exploit such agro waste for food production, especially in the era of food insecurity and move to actualize millennium development goals. Nutritional value of food around Umudike and Africa are posed with low vitamin capacity, which has resulted to vitamin deficiency issues among children. Since supplementation of mushroom has shown good correlation with high yield and nutritional richness (Okwulehie and Okoro, 2014), it therefore pertinent to try enriching the mushrooms with vitamins.

This fact i resulted to fortifying the substrates with vitamins since it has been established that the richness of mushrooms fruit bodies is a subject of the type of substrates on which they are grown.


1.1.6    Aims and Objectives

      The objective of this research work is:-

-      To determine the effect of straw substrates on the heavy metals composition of P. ostreatus

-      To determine the effect of straw substrates on the  minerals composition of P. ostreatus

-      To determine the effect of straw substrate on the phytochemical composition of P. ostreatus

 


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