PROJECT PROPOSAL ON

BIO-CONVERSION OF SOME SELECTED AGRICULTURAL

WASTE TO VALUE ADDED RUMINANT FEED WITH
Pluerotus florida AND Pluerotus ostreatus

BY

ALIU AHMED O.

F/HD/20/3294002

SUPERVISOR

DR. AKINFEMI A.

AUGUST, 2022
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 CHAPTER ONE

INTRODUCTION

1.1 Background of the Study

In recent days, ruminant animal in Nigeria suffer from under feeding especially during the dry
seasons due to shortage of forages and more so because of low nutritive value of available crop
wastes. The available wastes such as maize stover, corn cob, and maize husk cereal straws are
not able to meet the nutritional requirements of ruminants. To dispose these wastes, they are
usually burnt in heaps thereby releasing offensive odor and gases into the atmosphere. Some are
even thrown into the rivers and streams thereby endangering aquatic life (Jonathan et al, 2008).

Maize husks are fibrous leafy materials covering the maize ear. They are usually heaped in
refuse dump, farm lands and sometimes near homes. They usually constitute nuisance to the
environment and are rarely relished by ruminant animals because of their tough nature and low
nutritive value. Maize husk could be recycled and used as a source of valuable lignocellulosics
biomass for animals if treated with fungi.

Fahad et al., (2015), the bioconversion of agricultural residues into food has attracted the world
attention in recent years. Mushrooms are fungi that produce lignocellulosic enzymes, which
degrade complex organic matter and absorb the soluble substances (Ingale and Ramteke
2010).They can easily grow on almost all types of cellulosic residues such as banana leaves,
dried paddy straws, cotton waste and rice straws, sawdust enriched with poultry droppings
(Onuoha, 2014), Jatropha (Luiz et al., 2014) and even invasive weed species (Mintesnot et al.,
2014).

Oyster mushrooms are one of the most popular edible mushrooms and belong to the genus
Pleurotus and the family Pleurotaceae (Kong, 2004). The genus Pleurotus, is widely cultivated
due to its favorable organoleptic and medicinal properties, simple and low cost production
technology and higher biological efficiency (Chirinang and Intarapichet, 2009). Oyster
mushroom (Pleurotus spp.) is in the second position (after white button) in the world and its
cultivation has increased rapidly during the last decade. Oyster mushroom cultivation can play an
important role in managing organic residues whose disposal has become a problem (Lakshimi

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and Sornaraj 2014,Yu et al.,2014]. Furthermore, mushroom spent (grown media with mycelium)
can be used as an organic fertilizer or animal/fish feed (Fahad et al., 2015)

According to Akinfemi et al., (2009) cultivation of edible mushrooms like Pleurotus tuber-
regium, Pleurotus pulmonarius, Pleurotus sajor caju and Lentinus subnudus on lignocellulosic
wastes may thus be valuable for converting these farm waste materials, which are considered to
be waste into protein rich ruminant feeds. The cultivation and harvest of these fungi on maize
husk, apart from enriching the substrate, may also offer economic incentive for agribusiness.

Pleurotus florida is an edible mushroom having excellent flavor and taste (Mondal et al., 2010)
and is increasingly becoming popular as protein-rich delicious vegetable (Ng’etich et al. 2013,
Reyes et al. 2013). Pleurotus florida cultivation is popular due to low cost technology and easy
availability of various substrates for its cultivation. It is widespread in temperate, subtropical and
tropical zones and it shows the highest yield among the Pleurotus species.

Pleurotus ostreatus is a cultivated species of mushrooms characterizing with unique medical

properties. Its nutritional value comes from proteins, carbohydrates, fatty acids, vitamins and
mineral nutrients present in their fruit bodies (Salata et al., 2018). Active substances present in
the mushrooms have antioxidant, antibacterial**** white rot basidiomycete which belongs to the
subclass of ligninolytic microorganisms that produce laccases, manganese peroxidase, amylase,
cellulase, pectinase and protease (Abdou, 2003; Fan et al., 2008; Rashad et al., 2009). Many
researchers have studied chemical and biological pretreatments for enhancing conversion of
lignocellulosic materials to protein enriched product (Vijaya et al., 2007). Another alternative
method, using combined treatment with white rot fungi and yeast was developed; it is typically
used for bioconversion of those lignocellulosic wastes to protein enriched product (Galila et al.,
2012).

1.2 Objective of the Study

The aim of this study is to investigate the Bio conversion of maize husk into value added
ruminant feed with Pluerotus florida and Pluerotus ostreatus

The specific objectives of the study is:

i. To determine changes in the proximate composition and crude fibre fraction

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 ii. To determine the in vitro gas production for a period of 24hours
iii. To estimate the organic matter digestibility (OMD), short chain fatty acid (SCFA) and
metabolizable energy (ME) from the results of the proximate composition in vitro gas
production.

1.3 Justification of the Study

The cultivation of edible mushrooms has a great potential for the production of protein rich
quality food and for recycling of cellulosic agro- residues and other wastes and serves as the
most efficient and economically viable biotechnology for the conversion of lignocellulosic waste
materials to high quality food.

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 CHAPTER TWO

LITERATURE REVIEW

The following literature will be reviewed

2.1 Ruminant production in Nigeria

2.2 Feeding habit

2.3 Digestibility

2.3.1 Effect of Supplmentation on intake and Digestibility

2.4 Proximate Composition

2.4.1 Dry Matter Content

2.4.2 Moisture Content

2.4.3 Crude Fibre

2.4.4 Crude Protein

2.4.5 Ash

2.4.6 Ether Extract

2.4.7 Nitrogen Free Extract

2.5 Description of Maize

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 CHAPTER THREE

MATERIAL AND METHODS

3.1 Location of the Experiment

The experiment will be carried out in the laboratory of Federal University of Agriculture,
Abeokuta (FUNAAB) Ogun State.

3.2 Test Substrates

Dried samples of maize residues (maize husk) will be collected from the Teaching and Research
Farm, Yaba College of Technology, Nigeria. The materials were milled and oven-treated at 650C
until a constant weight was obtained for any dry matter determination.
The sporophores of Pleurotus florida (PF) and Pleurotus ostreatus (POT), growing in the wild
will be collected from Universityy of Ibadan, Oyo State, Nigeria. These will be tissue cultured to
obtain fungal mycelia (Jonathan and Fasidi, 2001) cited by Akinfemi e al. (2009). The pure
culture to be obtained will be maintained on plate of potato dextrose agar.

3.3 Experimental Designs

80g of dried milled sample of maize husk will be rationed to four treatments with four replicates
per treatment each with 5g

T1- Control Untreated maize husk

T2- maize husk treated with pleurotus florida MHTPF

T3- maize husk treated with pleurotus ostreatus MHTPOT

T2- maize husk treated with pleurotus florida and pleurotus ostreatus MHTPC

3.4 Inoculation

After sterilization, the samples will be inoculated at the centre each with P. florida and P.
ostreatus and combination of both of 2% mycelia disc and will be tied immediately. They will be
kept in the dark cupboard in the laboratory at 30 oC and 100% relative humidity (RH). After 21
days for PF, POT and the combination, the subststrates will be harvested by nautoclaving to

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terminate the mycelia and then then they will be oven dried to constant weight prior to chemical
analysis, in vitro gas production technique and in vitro digestibility

3.5 Chemical analytical methods

The proximate composition such as moisture content, ash, crude fiber, crude protein ether
extract, neutral and acid detergent fiber, micro and macro elements (Fe, Zn, Cu, Ca, P, K, Mg,
Mn, Pb, Ni and Cd) will be determined according to separate methods described in AOAC
(2012). Total hydrolyzable carbohydrates will be determined according to Montgomery (1961).
Lignocellulosic fractions based on dry matter basis will be determined according to the method
of Van Soest and Robertson (1980).

3.6 Statistical Analysis

Data will be collected using Analysis of Variance (ANOVA) and significant difference among
means will be compared usin Duncan Multiple Range Test (SAS, 2020) package at 5% level of
probability

The statistical model is shown below

YIJ= U + TI + EIJ

Where;
YIJ= Observed value of dependent variable
U= Population mean
TI = Mean effect of selected browse plants (forages)
EIJ= Random residual error

Conclusion

This study will be focused on treatments of maize husk as agricultural by product with P. florida
and P. ostreatus in order to increase crude protein and improve digestion in ruminant animals.
Furthermore, this study will open useful applied access for the production of maize stalk based
products of good quality. Moreover, the current study can add applied values based on the fungal
treatment of non-beneficial agricultural waste.

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 REFERENCES

Akinfemi A, Olaniyi J. B and Segun G.J. (2009) Bioconversion of maize husk into value added
ruminant feed by using white-rot fungus

AOAC, (2002). Official Methods of Analysis of Association of Official Agriculture Chemists

International 11th Ed., Washington.

Chirinang, P. and K. Intarapichet, K. (2009). Amino acids and antioxidant properties of the
oyster mushrooms, Pleurotus ostreatus and Pleurotus sajor-caju. Science Asia, 35: 326-
331.

Fahad, A., Ahmed, K., Ronnel, F. and Renato, G.R (2015).Cultivation of Oyster Mushroom
(Pleurotus florida) on Date Palm Residues in an Environmentally Controlled Conditions
in Saudi Arabia. Advances in Environmental Biology 9(3) 955-962

Galila A.M.A. Darwish, A.A. Bakr, M.M.F. Abdallah (2012) Nutritional value upgrading of
maize stalk by using Pleurotus ostreatus and Saccharomyces cerevisiae in solid state
fermentation. Annals of Agricultural Science 47-51.

Ingale, A. and A. Ramteke, A. (2010). Studies on cultivation and biological efficiency of

mushrooms grown on different agro-residues. Innovative Romanian Food Biotechnology,
6: 25-28.

Jonathan S. G.; I. O. Fasidi, A. O. Ajayi and O. Adegeye. (2008). Biodegradation of Nigerian

wood wastes by Pleurotus tuber regium (Fries) Singer. Bioresource Technology 99 (4):
807-811.

Kong, W. S., 2004. Descriptions of commercially important Pleurotus species. In: Mushroom
Growers’ Handbook 1, Oyster Mushroom Cultivation, Mushworld , Ch.4 pp:. 54-61

Lakshmi, S. S. and R. Sornaraj, R. (2014). Utilization of seafood processing wastes for

cultivation of the edible mushroom Pleurotus flabellatus. African Journal of
Biotechnology,. 13(17): 1779-1785.

8
Luz, J. M. R., M.D. da Nunes, M. D. S.A. Paes, S. A. D.P. Torres, D. P. and M.C.M. Kasuya, M.
C. M. (2014). Bio-detoxification of Jatropha curcas seed cake by Pleurotus ostreatus.
African Journal of Microbiology Research, 8(11): 1148-1156.

Mintesnot, B., A. Ayalew, A. and A. Kebede, A. (2014). Evaluation of biomass of some invasive
weed species as substrate for oyster mushroom (Pleurotus spp.) cultivation. Pakistan
Journal of Biological Sciences, 17 (2): 213-219.

Mondal, S. R., M.J. Rehana, M.S. M. J., Noman, M. S. and S.K. Adhikary, S. K. (2010).
Comparative study on growth and yield performance of oyster mushroom (Pleurotus
florida) on different substrates. Journal of Bangladesh Agricultural University, 8(2): 213-
220.

Montgomery, R., (1961). Further studies of phenol sulphuric acid reagent for carbohydrates.
Biochem. Biophys. Acta 84, 591–593.

Ng’etich, O.K., O.I. Nyamangyoku, J.J. O.I., Rono, A.N. J.J., Niyokuri, A.N. and J.C.
Izamuhaye,J.C. (2013). Relative performance of oyster mushroom (Pleurotus florida) on
agro-industrial and agricultural substrate. International Journal of Agronomy Plant
Production, 4(1): 109-116.

Onuoha, C. I., E.C. Nwachukwu E. C., I.O. Ezeibekwe I. O. and E.C. Nwagbara, E. C. (2014).
Evaluation of the effects of the combination of NPK fertilizer, cow dung, humus soil and
poultry droppings with sawdust on the number of days to primordia formation, maturity
and harvest of Pleurotus ostreatus. Journal of Yeast and Fungal Research,. 5(9): 103-
109.

Reyes, R.G., S.P. Kalaw, R.M.R. S.P., Dulay, H. R.M.R., Yoshimoto, N. H., Miyazawa, N. and
F. Eguchi, F. (2013). Philippine native and exotic species of edible mushrooms grown on
rice straw based formulation exhibit nutraceutical properties. Philippine Agricultural
Scientist,. 96(2): 198-204.

Van Soest, P.J., Robertson, J.B., (1980). In: Standardization of Analytical Methodology for
Feeds. IDRC Publ. 134 e, International Development Research Center, Ottawa, pp. 49–
60.

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Yu, H.L., B.B. Ly B.B., Q. Tan, X.D. Q., Shang, C.Y. X.D., Song, L.J. C.Y., Zhang, M.Y. L.J.,
Zhang, M.Y. and G.X. Li, G.X. (2014). Cultivation of Pleurotus cornucopiae using
biogas production residue. Acta Edulis Fungi,. 21(1): 25-28.

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