PRODUCTIVITY AND BIO-SAFETY ASSESSMENT OF AQUEOUS EXTRACT OF PLEUROTUS OSTREATUS (JACQ.EX.FR) P. KUMM. FRUIT-BODIES CULTIVATED ON ACID-INDUCED SUBSTRATE

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ABSTRACT


This investigation was conducted to study the productivity and bio-safety of Pleurotus ostreatus fruit bodies cultivated on Hydrochloric acid (HCl)-induced oil palm bunch (OPB) substrate. Percentage solutions (0.1-0.5%) HCl were used to induce changes on the initial pH (10.1) of 1400g of OPB replicated 7 times; to 8.2, 7.8, 7.4, 7.1, 6.1 and control (9.0) respectively. One way analysis of variance (ANOVA) was adopted for data analysis using SPSS version 20.1. Means were separated by Duncan Multiple Range Test (DMRT) at p≤0.05%. Fruiting was delayed (19days) in 0.5%HCl substrate while control fruited after12 days. 24 Wistar albino rats made into six groups of four (4) replications, were administered 500ml/kg aqueous extract of mushroom at 24hr interval, for 21days. 0.5% substrate induced the highest (1799.10g/kg) fruit body yield, number (705) and biological efficiency (137.97%) while control produced the lowest (865g/kg, 424 and 61.79.3%). Fruit bodies from 0.2% HCl OPB substrate had the highest mean cap diameter (7.75±0.29cm), stipe length (3.00±0.12cm) and Weight (9.02±0.79g) while those of 0.1% (5.96±0.23cm), 0.4% (2.44±0.07cm) and 0.1% (5.23±0.52g) had the lowest. Fruit bodies from 0.4% had the highest (16.72±0.57mg/100g) vitamin B1 content among other levels while the lowest concentrations of vitamins A and E were observed at all levels. Phenolics were highest at 0.1% (188.96±6.01%) but lowest at (0.2% (106.56±1.76%). Tannins increased from 101.06% in fruit bodies from 0.2% - 134.68 in those of 0.5% substrate groups, unlike other levels. Alkaloids were highest at 0.2% (46.87±0.43%) and terpenoids at control. Moisture (9.07±0.15%) and Protein (24.98±0.03%) contents were highest at 0.5%HCl while control (80.09±0.09%) had the highest carbohydrate. Ash, ether extract (E.E) and crude fiber (CF) were low across all treatment levels and control. Sodium (149.85±6.32) and potassium (5.07±0.46mg/100g) were highest in mushrooms of 0.5%HCl while Chlorine and Calcium tend to increase with increase in percentage HCl. Concentrations of Zinc (181.07±1.22mg/100g) and Iron (197.70±20.01mg/100g) were highest in 0.1% HCl while those of Selenium, Lead and Copper were among the lowest. No mortality was recorded among the animals used. Rats of 0.2 (690.00±13.54 ×109/L) and 0.3% (13800.00±115.47×109/L) groups had the highest white and red blood cells. Highest platelets count of 237.50±23.94 ×109/L was recorded in the control, 0.1 and 0.5%HCl groups. 0.1, 0.4 and 0.5% groups had the highest level of haemoglobin while 0.4(42.25±1.32%) and 0.5% (42.00±0.82%) groups had the highest packed cell volume. Liver function test showed that 0.3HCl group had the highest aspartate aminotransferase AST (37.08±1.36IU/L) and alkaline phosphatase (ALT) (29.71±3.68IU/L) levels while control had the highest alanine aminotransferase (ALP) (23.90±1.15 IU/L). Albumin and Bilurubin levels were not affected by the mushroom extract while Urea and serum protein levels of 0.1 (35.83±1.31 mmol/L) and 0.3%HCl (6.00±0.16 mmol/L) groups had the highest. GSH, GPx, MDA, SOD and catalase were not significantly affected at all levels (0.1-0.5%) groups. Both In vitro and In vivo bio-safety studies were not significant (p ≥ 0.5), suggesting that the mushrooms could be safe for human consumption. Oil palm bunch should therefore be adopted in the commercial production of the Oyster mushroom.






TABLE OF CONTENTS


Title Page                                                                                                                    i

Declaration                                                                                                                 ii

Certification                                                                                                               iii

Dedication                                                                                                                   iv

Acknowledgements                                                                                                    v

Table of Contents                                                                                                       vi

List of Tables                                                                                                              xi

List of Figures                                                                                                                                     xii

List of Plates                                                                                                                                       xiii

Abstract                                                                                                                                             xiv

 

CHAPTER 1: INTRODUCTION

1.1       Life Cycle of Mushroom                                                                                1

1.2       Taxonomy                                                                                                       3

1.3       Economic Importance of Oyster Mushrooms                                                 4

1.3.1    Nutritional and food values                                                                            4

1.3.2    Medicine/tonic                                                                                               6

1.3.3    Income and job creation                                                                                 6

1.4       Justification of Study                                                                                      7

1.5       Statement of the Problem                                                                               8

1.6       Specific Objectives                                                                                         9

 

CHAPTER 2: LITERATURE REVIEW

2.1       Mushroom and Food Security                                                                        11

2.2       Effect of pH on Mushroom Fruit Body Formation                                        12

2.3       Substrates used in Mushroom Cultivation                                                     13

2.4       World Mushroom Production                                                                         17

2.5       World Mushroom Market                                                                               21

2.6       Important Mycochemicals in Mushroom                                                      22                                                        

2.7          Mushroom Toxicity                                                                                     25

2.8          Toxic Effect of Poisonous Mushrooms                                                       27

2.9          Case Reports on Mushroom Poisoning                                                       33

2.10        Heavy Metal Uptake by Mushrooms                                                           38

2.11        Bioaccumulation in Mushrooms                                                                  39

2.12        Tissue and Organ Responses to Toxins                                                       39

2.12.1     Effect of toxic substances on the liver                                                        39

2.12.2     Toxic effects of plants on the liver                                                              43

2.12.3     Effect of toxic substances on the kidneys                                                   45

2.12.4     Alkaline phosphatase (ALP) concentration                                                49

2.12.5     Albumin concentration                                                                                50

2.12.6     Uric acid concentration                                                                               50

2.12.7     Urinary protein                                                                                            51

2.12.8     Hematological parameters and relationship with diseases                          51

2.12.8.1  Blood cells                                                                                                   51

2.12.8. 2 Red blood cells (RBC)                                                                                 52

2.12.8.3  White blood cells (WBC)/leucocytes                                                          54

2.12.8.4  Pack cell volume (PCV)                                                                              57

2.12.8.5  Packed cell volume (PCV) hemoglobin (Hb) levels                                   57

2.11.8.6  Platelets count                                                                                             58

 

CHAPTER 3: MATERIALS AND METHODS

3.1          Study Area                                                                                                  59

3.1.1       Mushroom cultivation                                                                                 59

3.2          Source of Spawn Culture                                                                          59

3.2.1       Spawn production/multiplication                                                                59

3.2.2       Substrate preparation                                                                                   60

3.3          Determination of Substrate pH                                                                    60

3.4          Experimental Design                                                                                   60

3.5          Substrate Inoculation                                                                                   60

3.6          Measurement of Morphological Characters                                                            61

3.6.1       Stipe size of fruit bodies                                                                              61

3.6.2       Cap diameter                                                                                               61

3.6.3       Effect of substrates on fruit body number of the mushrooms                   61

3.7          Yield and Biological Efficiency                                                                  61

3.8          Sample Preparation                                                                                     62

3.9          In Vitro Assay                                                                                              62

3.9.1       Proximate analysis                                                                                       62

3.9.1.1    Determination of crude protein                                                                   62

3.9.1.2    Determination of moisture content                                                              62

3.9.1.3    Determination of ash contents                                                                     63

3.9.1.4    Determination of carbohydrate (CHO)                                                        63

3.9.1.5    Determination of ether extract                                                                    63

3.9.1.6    Determination of crude fibre                                                                       64

3.9.2       Determination of vitamins                                                                           64

3.9.2.1    Determination of vitamin A (retinol)                                                          64

3.9.2.2    Determination of vitamin B1 (thiamin)                                                       65

3.9.2.3    Determination of vitamin B3 (niacin)                                                          65

3.9.2.4    Determination of vitamin C (ascorbic acid)                                                            66

3.9.2.5    Determination of vitamin K (phylloquinone)                                             66

3.9.2.6    Determination of vitamin E (tococpherol)                                                  67

3.9.3       Determination of percentage bioactive compounds                                                68

3.9.3.1    Determination of phenolics content                                                            68

3.9.3.2    Determination of tannins                                                                             69

3.9.3.3    Determination of sterols                                                                              69

3.9.3.4    Determination of alkaloids                                                                          70

3.9.3.5    Determination of terpenoids                                                                        70

3.9.3.6    Determination of glycosides                                                                        70

3.9.4       Determination of minerals                                                                           71

3.9.5       Determination of heavy metals                                                                   72

3.10        In Vivo Assay                                                                                               72

3.10.1     Preparation of aqueous extract of P. ostratus                                              72

3.10.2     Experimental animals                                                                                  72

3.10.3     Determination of LD50                                                                                                                         73

3.10.4     Grouping of animals                                                                                    74

3.10.5     Measurement of body weight                                                                      74

3.10.6     Administration of the extract                                                                      74

3.10.7     Preparation of animals for sacrifice                                                            74

3.10.8     Determination of haematological parameters                                             74

3.10.8.1  Red blood cell (RBC)/erythrocyte count (x109/L)                                       74

3.10.8.2  White blood cell (WBC)/ leucocyte count (×109/L)                                   75

3.10.8.3  Platelets count (x109/L)                                                                               76

3.10.8.4  Determination of packed cell volume (PCV) (%).                                      77

3.10.9      Liver functioning test                                                                                 77

3.10.9.1  Determination of aspartate aminotransferase (AST) (IU/L)                      77

3.10.9.2  Determination of serum alanine aminotransferase (ALT) activity (IU/L)  77

3.10.9.3  Determination of serum alkaline phosphatase (ALP) activity (IU/L)  78

3.10.9.4  Determination of serum bilirubin                                                                79

3.10.9.5  Determination of serum albumin                                                                80

3.10.9.6   Determination of serum total protein                                                         80

3.10.10    Kidney tests                                                                                                81

3.10.10.1 Determination of urea (mmol/L)                                                                81

3.10.11    Determination of enzymatic antioxidants and catalase                              83

3.10.11.1 Estimation of reduced glutathione (GSH)                                                  83

3.10.11.2 Estimation of glutathione peroxidase (GPx)                                              84

3.10.11.3 Estimation of extent of lipid peroxidation (malondialdehyde/MDA)       85

3.10.11.4 Estimation of superoxide dismutase (SOD)                                               85

3.10.11.5 Estimation of catalase                                                                                86

3.11         Statistical Analysis                                                                                     87

 

CHAPTER 4: RESULTS AND DISCUSSION

4.1          Results                                                                                                         88

4.2          Discussion                                                                                                    118

4.2.1       pH variations in substrate and formation of mushroom fruit bodies  118

4.2.2       Morphological characteristics of fruit bodies                                              119

4.2.3       Productivity and biological efficiency of P. ostreatus fruit bodies                        120

4.2.4       In vitro assay                                                                                                121

4.2.4.1    Vitamins concentrated (mg/100g) of P. ostreatus fruit bodies                        121

4.2.4.2    Bioactive compounds concentration of the fruit bodies                              122

4.2.4.3    Proximate composition of fruit bodies                                                        124

4.2.4.4    Minerals concentration of fruit bodies                                                        125

4.2.4.5    Heavy metals concentration of fruit bodies                                                 126

4.2.5       In vivo assay                                                                                                128

4.2.5.1    Acute toxicity test                                                                                        128

4.2.5.2    Effect of mushroom extract on the hematological parameters                        128

4.2.5.3    Effect of mushroom extract on the liver                                                      131

4.2.5.4    Serum protein concentration                                                                       134

4.2.5.4    Effect of mushroom extract on kidney function                                          134

4.2.5.4.1 Serum urea concentration                                                                            134

4.2.5.5    Effect of mushroom extracts on antioxidant enzymes and catalase  135

 

CHAPTER 5: CONCLUSION AND RECOMMENDATIONS

5.1          Conclusion                                                                                         138

5.2         Recommendations                                                                                        139    

              References                                                                                                    141

              Appendices                                                                                                   165

 

 





LIST OF TABLES

 

1.1.      Some recognized mushroom toxins with specific/deadly effects                  32

4.1.      pH of substrates and fruiting duration of P. ostreatus                             88

4.2.      Morphological characters of fruit bodies                                                 89

4.3.      Productivity and biological efficiency of fruit bodies                              90

4.3.1.     Vitamins concentration (mg/100g) of fruit bodies                                         91

4.4.      Phytochemical (%) compounds composition of fruit bodies                   93

4.5.      Proximate composition (%) of fruit bodies                                              95

4.6.      Minerals concentration (mg/100g) of fruit bodies                                   97

4.7.      Heavy metals concentration (mg/100g) of fruit bodies                            98

 


 

 

 

 

 

 

LIST OF FIGURES

4.1.         Effect of HCl on the WBC count in albino rats                                            100

4.2.          Effect of HCl on the packed cell volume (PCV) level                                  101

4.3       Effect of HCl on the red blood cell (RBC) level                                            102

4.4.      Effect of HCl on the haemoglobin (HB) levels                                              103

4.5.      Effect of HCl concentration on the platelets level                                         104

4.6.      Effect of HCl on the AST levels in the liver                                                  105

4.7.      Effect of HCl on the alanine aminotransfarase (ALT) level                          106

4.8.      Effect of HCl on the ALP levels in the liver.                                                 107

 4.9.     Effect of HCl on the albumin level                                                                108

4.10.    Effect of HCl on the T. bilirubin Level                                                          109

4.11.    Effect of HCl on the D. bilirubin Level                                                          110

4.12.    Effect of HCl on the urea level in the kidney.                                                111

4.13.    Effect of HCl on the serum protein level.                                                      112

4.14.    Effect of HCl on the glutathione (GSH) level                                                113

4.15.    Effect of HCl on the glutathione peroxidase (GPx) level.                             114

4.16.    Effect of HCl on the MDA levels                                                                   115

4.17.    Effect of HCl on the superoxide dismutase (SOD) level                               116

4.18.    Effect of HCl on the catalase level                                                                 117

 

 

 

 

 

 

 

 

 

 

LIST OF PLATES

 

1          Uncolonized spawn substrate                                                                         163

2          Fully colonized spaw                                                                                      163

3          OPB substrate                                                                                                 163

4          Fruit bodies from different groups                                                                 163

5          C. cinerius contaminated OPB                                                                       164

6          Ground fruit bodies                                                                                        164

7          Dried fruit bodies                                                                                            164

8          Collection of samples from rats                                                                      164     

 

 






 

CHAPTER 1

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 pre-formed food like smaller broken down molecules of lignin, cellulose and starch (Banjo, 1998). Chang, (1999) 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 unaided eye and to be picked up by hand. Ideally, the word mushroom refers only to the fruit body of a macro-fungus. Unlike green plants, mushrooms are heterotrophs and without chlorophyll, they cannot generate nutrients by photosynthesis, but instead take already made nutrients from organic materials. Most mushroom species are either under the Basidiomycota or Ascomycota; the two phyla are under the kingdom Fungi (Cho, 2004).

 

 1.1      LIFE CYCLE OF MUSHROOM

The life cycle of a mushroom may be traced from a spore, which under favorable condition germinates to form a mass of branched hyphae of mycelium which colonizes a substrate. This represents the vegetative stage of its growth. When a given substrate is fully colonized, the vegetative growth ceases. Typically some hyphae form primordia or fundament which is the beginning of the reproductive stage (Bahl, 1985). This develops further to differentiate into stipe (stalk) and the pileus (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 (Takama et al., 2010). 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 parts of the world including Nigeria. The growing and consumption interest of oyster mushroom is increasing largely due to its taste, medicinal and nutritional properties (Garcha et al., 1993).

 

Oysters are naturally found on rotten wood materials. Takama et al., (2010­) observed that this fungus is common in Nigeria and often found growing around the African breadfruit tree (Treculia africana). In Nigeria, the most prized edible species are Pleurotus, Termitomyces, Tricholoma and Volvariella (Ambali et al., 2008).

 

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, 1999). Oyster mushrooms are the third largest commercially produced mushroom in the world (Obodai et al., 2003); 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., 2014).

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 acceptable edible biomass of high market value.

 

1.2       TAXONOMY

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

Kingdom         Mycota

Division          Basidiomycota

Class               Agaricomycetes

Order               Agaricales

Family             Pleurotaceae

Genus              Pleurotus

Species            P. ostreatus (Jacq.Ex.Fr) P. Kumm

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), was proposed as a genus by Quelet, (1886). Three genera of this group, Pleurotus, Lentinus, and Panus, were possible to be separated according to the 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.


1.3       ECONOMIC IMPORTANCE OF OYSTER MUSHROOMS

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

1.3.1    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 which 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 flavour, they also offer themselves as potential protein, minerals and vitamins 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 et al., 1982; Furlani, 2014). 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 Okogbo, (1991), Okwulehie and Odunze, (2004a), mushrooms generally contain low oil and fat, and are therefore 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 a misdirection (Aletor, 1995). Fasidi and Kadiri, (1990), 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 Okogbo, 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.

1.3.2    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). Some kinds of mushrooms such as Amanita muscaria had been reported to produce intense excitement and hallucination to the consumer (Kandler, 1983; Oei, 1996; Royse, 1996). Mushroom has also been reported as therapeutic food useful in preventing diseases such as hypertension, hyper-cholesterolemia and cancer. These functional characteristics are mainly due to their chemical composition (Manzi, 1999b). Owing to the tolerance of P. pulmonarius to high temperature, it has been reported that its speedy fruiting and yield efficiencies attract many cultivators to the mushroom industry (Trudell and Ammirati, 2009).  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 as anti-tumour, anti-cancer and for many other therapeutic purposes (Liu et al., 2001; Chang and Miles, 2004).

 

1.3.3    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).


1.4       JUSTIFICATION OF STUDY

Using modern approaches, scientists have isolated and identified from mushroom, specific components that can either destroy or at least debilitate three of mankind killer diseases: cancer, heart disease and HIV/AIDS (Tricita, 2004). Research has also proved that some mushroom species are highly medicinal and are found to reduce cholesterol, inhibit tumor counteract Pernicious anaemia etc (Rambelli and Menini, 1983; Stamets, 1993; Oei, 2003). Apart from their potentials for medicines, mushrooms are also good source of protein, minerals and vitamins (Walid et al., 1988).

Available records show that mushroom industry is a multi-billion Dollar enterprise in many countries of the world especially, the developed nations. This has also created and provided employment to out-of-school youths and even to school children (Oei, 2003; Tricita, 2004).In spite of this global trend in mushroom production, most edible mushrooms are still being collected from the wild in many localities in Nigeria.

Oil palm bunch (OPB) is one of the most abundant organic materials often seen as waste around homes and palm oil mills. Even though, people living in most rural communities have used it to produce soap of very low market quality, other importance of it are yet to be fully harnessed. The incorporation of oil palm bunch in mushroom cultivation and other waste management strategies will no doubt boost food security and employment opportunities.

 

1.5   STATEMENT OF THE PROBLEM

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). The ongoing awareness on mushroom as a valuable source of protein with low cholesterol content which over rides meat and other fatty foods, may soon diminish due to the fore-scarcity of sawdust which serves as major substrate for its commercial production (Quimio et al., 1990; Jonathan et al., 2012c).

 

Oil palm bunch has reportedly contributed high level environmental degradation especially around oil palm mills and other areas where palm oil is commercially processed. To safeguard our environment, oil palm bunch must be put to adequate use, either by transforming it into alternative economically important product or through biological degradation which mushroom is capable of doing.

 

However, 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 competitor mushroom that contaminates mushroom bed) 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 a large scale production, Oyster mushroom growers prefer to use lime (CaCO3) as an alkaline buffer to optimize pH of some acidic substrates to increase mushroom production and maximize profit, but a suitable acid buffer to optimize the pH of highly alkaline substrates such as OPB has not been well understood by experts.

Many questions have been raised on the consumption safety of P. ostreatus if grown on Hydrochloric acid (HCl) induced-OPB as a means of optimizing its pH for better yield of mushrooms; since  Mycophobia (ie, irrational fear of consuming fungi such as mushrooms) is a major problem facing mushroom consumption in Nigeria (Kalu et al., 2013).

In Nigeria, proper documentation of mushroom toxicity is rare, as most reports on toxicity or death in humans associated with mushroom consumption are under reported. Besides, several new syndromes are being described in mushroom poisoning (Diaz, 2005), which calls for their continuous toxicological evaluation.

In view of possible toxicity of  P. ostreatus fruit bodies grown on HCl mediated Oil Palm Bunch (OPB), the need for both in vitro and in vivo toxicological screening of the mushrooms become pertinent.


1.6       SPECIFIC OBJECTIVES

The main objectives of this experiment are to:

i.      Determine the effect of HCl on the pH of oil palm bunch (OPB)

ii.     Determine the yield and Biological efficiency of P. ostreatus cultivated on HCl induced OPB.

iii.   Evaluate the effect of HCl on the morphological characteristics of P. ostreatus fruit bodies.

iv.   Determine the nutritional, vitamins, phytochemicals and heavy metals composition of the mushroom fruit bodies.

v.     Assess the in vivo bio-safety of the P. ostreatus fruit bodies

 


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