MICHAEL OKPARA UNIVERSITY OF AGRICULTURE UMUDIKE,

UMUAHIA, ABIA STATE.

A TECHNICAL REPORT ON

THE STUDENT INDUSTRIAL WORK EXPERIENCE PROGRAMME

CARRIED OUT AT

THE FEDERAL INSTITUTE OF INDUSTRIAL RESEARCH, OSHODI LAGOS,

BY

……….

(MOUAU/)

DEPARTMENT OF BIOCHEMISTRY

COLLEGE OF NATURAL SCIENCES.

IN PARTIAL FULFILLMENT OF THE REQUIREMENTS OF THE COURSE BCH399

STUDENTS’ INDUSTRIAL WORK EXPERIENCE SCHEME (SIWES)

FOR THE AWARD OF BACHELOR OF SCIENCES (B.Sc.) HONS. BIOCHEMISTRY.

SEPTEMBER, 2025
 DEDICATION

This report is dedicated to God Almighty, who in His in infinite mercies kept me and made

my SIWES program a huge success.

 ACKNOWLEDGEMENT.

My profound gratitude is to the Almighty God who has kept me in good health and given me

wisdom to go through this training. I appreciate the efforts and support of my loving parents.

Also Dr. Frank Anayo Orji and all biotechnology staff whom were of immense help during the

period. My multitude of friends, God bless you all.

 TABLE OF CONTENTS

Pages:

Title page i

Dedication ii

Acknowledgment iii

Table of contents iv

CHAPTER ONE

Introduction -------------------------------- 1

History of SIWES -------------------------------- 2

Aims &Objectives of SIWES ------------------------------ 3

CHAPTER TWO

History of FIIRO --------------------------------- 4

Mission, Vision & Objectives of FIIRO -------------------------------- 5

Organizational Chart of the Institute ------------------------------- 6

CHAPTER THREE

Experience gained during the training -------------------------------------- 12

Equipments/instruments found in the laboratory ------------------------------- 12

Biotechnology Unit: Cellulase enzyme production ------------------------ 13

Biotechnology Unit: Protease enzyme production -------------------------- 19

CHAPTER FOUR

Waste utilization and Fermentation Unit: Mushroom cultivation -------------- 26

Waste utilization and Fermentation Unit: Ethanol production ------------------ 33

CHAPTER FIVE

Challenges encountered -------------------------------------------------------- 40

Recommendation -------------------------------------------------------- 40
 CHAPTER ONE.

INTRODUCTION.

This report provides a rundown of the training I received at the Federal Institute of Industrial

Research, Oshodi during the course of the SIWES program. This report also covers the history

of SIWES, its aims and objectives, the challenges i encountered and possible solutions.

HISTORY/OVERVIEW OF THE STUDENT INDUSTRIAL WORK EXPERIENCE

SCHEME

The students industrial work experience scheme SIWES was established in the year 1973 by

the Industrial Training Fund (ITF) to solve the problem of lack of adequate practical skills

preparatory for employment into industries by Nigeria tertiary institution graduates.

The scheme educates students on industrial based skills essential for smooth transition from

the classroom to the working environment.

Students of tertiary institutions are given the opportunity of being familiarized and exposed to

the needed experience in handling machinery and equipment which are usually not available

in the educational institutions.

Partaking in the SIWES industrial training exercise has infact become a crucial pre-requisite

for the award of diploma and degree certificates in specific disciplines in most institutions of

higher learning in Nigeria in line with Act No. 47 of 1971 government education polices.
 This is an effort which was created in order to bridge the existing gap between the theory

taught in the classrooms and practice of science, agriculture, medicine, engineering,

technology and other professional programmes in the Nigerian tertiary institutions

The scheme is a tripartite programme, involving the students, the universities, and the

industry (employers of labour).

It is funded by the federal government of Nigeria and jointly coordinated by the Industrial

Training Fund (ITF) and the National Universities Commission (NUC), the National Board

for Technical education (NBTE) and National Communication for College of Education

(NCCE).

This is an effort to bridge the gap existing between the theory taught in the classrooms and

practice of agriculture, medicine, engineering, technology and other professional educational

programmes in the Nigerian tertiary institutions.

1.1 AIMS & OBJECTIVES OF SIWES

Specifically, the objective of the Students Industrial Works Experience Scheme are to:

 Provide an avenue for students in the Nigerian Universities to acquire industrial skills

and experience in their various course of study.

 Prepare students for the work situation they are likely to meet after graduation.

 Expose students to work methods and techniques in handling equipments and

machinery that may not be available in the universities.

 Make the transition from the university to the world of work easier, and thus enhance

students’contacts for latter job placements.

 Provide students with an opportunity to apply their theoretical knowledge in real work

situations, thereby bridging the gap between theory and actual practice.

 Enlist and strengthen employer’s involvement in the entire educational process of

preparing university graduates for employment in industry.

 CHAPTER TWO

2.0 HISTORY/ OVERVIEW OF THE FEDERAL INSTITUTE OF INDUSTRIAL

RESEARCH OSHODI(FIIRO)

The Federal Institute of Industrial Research, Oshodi (FIIRO) with the motto “Industrialization

through research” was established in 1956 and is currently headed by Prof. (Mrs.) Gloria

Elemo. The institute is a parastatal under the Federal Ministry of Science and Technology. It

came to be as a result of the idea of an economic mission sent to Nigeria in 1953 by the World

Bank due to the observation that industrial research activities in the nation were diffused and

uncoordinated with no particular direction (FIIRO, 2016). Currently, the areas of focus of the

institute include: Research & development of food and Agro-allied processing technologies,

Research & development into pulp and paper processing, Research & development into

packaging and product design and finally the design and fabrication of equipment prototype.

2.1 MISSION, VISION AND OBJECTIVES OF FIIRO

The mission, vision and objectives for which the institute was created are resonated in its

mission and vision statements.

2.1.1 VISION STATEMENT

To be the foremost Centre for Science and Technology based research and development for the

industrialization and socio-economic advancement of the nation.

MISSION STATEMENT

To conduct and promote market-driven research and development (R&D) for the

industrialization and socio- economic development of the nation.

OBJECTIVES OF FIIRO

The major objective of the institute is to assist in accelerating the industrialization of the

Nigerian economy by finding utilization for the country’s raw materials and also upgrading

indigenous production technologies. Specifically, the objectives of FIIRO include;

 Identifying and characterizing local raw materials for use in industries.

 To develop appropriate technologies as well as upgrade indigenous technologies in the area of

food and agro- allied processing and in various nonfood use.

 To develop pilot scale operations.

 To assist in the transfer, adaptation and utilization of these technologies by local enterprises.

 To undertake economic evaluation of projects and consultancy services.

The institution is divided several departments all of which play a role in the fulfillment of the

objectives of the institute. The research departments include: Biotechnology, Chemical, Fibre

and Environmental Technology (CFET), Food and Analytical services, Project design and

development (PDD), Planning, technology transfer and information technology (PTTIM),

Administration and supply (FIIRO, 2011).

FIIRO ORGANOGRAM:
 CHAPTER THREE

EXPERIENCE GAINED/ WORK DONE DURING THE INDUSTRIAL TRAINING

Upon resumption at FIIRO, I was posted to the Enzyme technology division of

Biotechnology department (BIOTECH) which is divided into the waste utilization and

fermentation laboratory, food safety and quality management system laboratory, molecular

biology laboratory, and Enzyme technology laboratory. However, I was opportuned to have

access to other divisions and departments within the organization; hence the wealth of

experience I gained in the course of the training is drawn from these departments.

INTRODUCTION TO THE DIFFERENT LABORATORY EQUIPMENTS

1. Weighing balance
2. Magnetic Stirrer-Hotplate
3. Autoclave
4. Drying oven
5. Hot-air oven
6. Lamina-air flow Chamber
7. Incubator
8. Microscope
9. Gyratory shaker incubator
10. Centrifuge
11. PH Meter
12. Membrane filtration Unit
13. Water bath
14. Spectrophotometer
15. Refrigerator
16. Freezer
17. Anaerobic jar
18. Bunsen burner
19. Refractometer
BIOTECHNOLOGY UNIT: ENZYME TECHNOLOGY DIVISION

Biotechnology unit specializes in the use of microorganisms or part of microorganisms to improve

and produce foods, beverages and chemicals such as enzymes, wines, alcohol, organic acids and

glucose syrup. Attention is paid to the utilization of industrial and domestic wastes to produce goods

and services

I was able to learn a number of things from this division. Some of which are detailed in this

section.

Enzyme Production;

This was carried out at the biotechnology department. I was privileged to participate in

the production and subsequent assay of cellulase and protease enzymes. The cellulase enzyme

was produced via the solid state fermentation method while the enzyme Protease was produced

via the Submerged fermentation method. The processes involved in the production and assay

for these two enzymes are detailed in this section.

Solid state fermentation production of CELLULASE enzyme using Brewer

spent grain by Aspergillus flavus.

Cellulase refers to the group of enzymes that act serially or synergistically to

decompose cellulosic material by catalyzing the hydrolysis of 𝛽 − 1,4 − linkages in Cellulose.

Cellulases breakdown the cellulose molecule into monosaccharides (simple sugars) such as

beta-glucose, or shorter polysaccharides and oligosaccharides. Cellulases contain non-catalytic

carbohydrate binding molecules (CBMs) and\or other functionally known or unknown

modules.

In nature, complete cellulose hydrolysis is mediated by a combination of three main types of

cellulases: (i). endoglucanases (EC 3.2.1.4), (ii). Exoglucanases, including cellobiohydrolases

The substrate brewer spent grain comprises of barley, sorghum, malt, wastes which contain
high level plant fibre (cellulose).

Substrates like wheat bran, sugar cane bagasse, tiger nut chaff, etc could also be used for the
production of the enzyme Cellulase.

Trichoderma ressei is also known to be a producer of cellulase enzyme extracellularly and can
be used in place of or with Aspergillus flavus.

Materials Used:

1. KH2PO4

2. 0.5M Citrate buffer at pH 6 for enzyme extraction.

3. MgSO4.H2O

4. 0.5M Citrate buffer at pH 4.8 for enzyme assay.

5. CaCl2.H2O

6. 3,5 dinitrosalicyclic acid.

7. FeSO4.7H2O

8. ZnSO4.7H2O

9. CoCl2.H2O

10. KNO3

11. Test tubes and tube rack.

12. Distilled water

13. Conical Flask

14. Autoclave

15. Cellulose powder

16. 1% Carboxyl methylcellulose solution

17. Aspergillus flavus spores

18. Water bath

 19. Grounded Filter paper

20. pH meter

21. Fermentation trays.

22. Spectrophotometer

23. Muslin cloth.

24. Brewer spent grain.

25. Tween 80 solution.

Procedure:

1. Specified quantities of KH2PO4, MgSO4.H2O, CaCl2.H2O, FeSO4.7H2O, ZnSO4.7H2O,

CoCl2.H2O, and KNO3 were measured into a 1000ml conical flask and dissolved with 200ml

of distilled water, The solution was made up to the 1000ml mark by adding more distilled

water.

2. The 1000ml mineral solution was now placed on a magnetic hotplate stirrer for homogenization

by a magnetic spindle bar.

3. About 500ml of the mineral solution is mixed with pre-weighed 500 grams Brewer spent grain

and mixed using clean hands. The resulting mixture is placed in a muslin bad and autoclaved

at 121°C for 45 minutes.

4. 300 ml of distilled water is weighed out into another conical flask and about 2ml of Tween 80

solution is added to it. The resulting mixture is autoclaved at 121°C for 15 minutes.

5. Fermentation trays are washed and swabbed with cotton wool and ethanol after which they

were wrapped with aluminium foil and placed in a hot air oven for sterilization at 165°C for

180 minutes (3 hours).

6. The plates containing fresh Aspergillus flavus isolates is gotten, little amount of the tween 80

mixture is poured on the surface of the culture and with the aid of a flamed wire loop the spores

are scraped off the surface and poured into the bulk tween 80 mixture.
7. The sterile BSG is poured into the sterilized tray and about 50 grams of cellulose powder is

added to it and mixed thoroughly.

8. 200 ml of the tween 80 solution containing the isolate spores is drawn and added to the BSG

mixture aseptically. The spores and the BSG are mixed with gloved hands.

9. The fermentation tray containing the whole mixture is covered with aluminium foil and

incubated at 25°C for 5-7 days.

10. After 7 days, the fermented BSG is aseptically turned into a clean bucket and a little portion of

the sodium citrate buffer is added to it, with gloved hands the buffer and the BSG mixture is

homogenized well first, then the rest of the buffer solution is turned in and mixed properly. The

whole mixture is covered and allowed to stand for atleast 40 minutes.

11. After 40 minutes, a clean container (conical flask) is gotten and a funnel is placed in it, a sterile

muslin cloth is placed in the funnel and the BSG and buffer mixture is filtered giving a dark

chocolate coloured liquid filterate. The filterate gotten is the crude cellulase enzyme ie. It still

contains a lot of impurities and has to first undergo centrifugation to obtain a clear solution and

then so many other purification techniques to get a pure enzyme.

12. Following extraction, the level of activity of the enzyme was assayed for. The substrates used

were grinded filter paper and 1% CMC solution. The details of the assay are summarized in

the table below as well as in the text following;

Table 1: Table showing some steps involved in the assay for cellulase.

Test tube Grinded CMC Citrate Produced

no filter paper solution(ml) buffer (ml) cellulase

(g) enzyme(ml)

1. 0.025 - 3.00 0.5

 2. 0.025 - 3.00 0.5

3. - 0.05 3.00 0.5

4. - 0.05 3.00 0.5

5. - - 3.5 -

All five test tubes with their various contents were incubated for 1hr in a water bath set at 50°C.

Afterwards, 3ml of DNSA was added to the content of each test tube. The test tubes and their

contents were then placed for 10 mins in a water bath at 100°C in order to terminate the

reaction.

13. Color changes were observed in the contents of the test tubes with the exception of that in

test tube 5 (blank).

14. The absorbance of each of the solutions was read in the spectrophotometer at a
wavelength of 540nm.

Figure 1: Depiction of assay. Picture on the left shows the DNSA addition stage ie; before
DNSa was added while that on the right shows the result obtained upon addition of DNSa
solution to terminate the whole reaction.
Conclusion; The activity of the enzyme now is determined by how much wavelength each of

the solutions in the various tubes can absorb or scater. The cellulase enzyme should be able to

degrade the cellulose materials in the test tubes to yield other simpler compounds which in

fact should increase the absorbance of the solution.

Submerged fermentation production of PROTEASE enzyme by Bacillus

subtilis, Bacillus polymyxa and Bacillus lichenoformis.

Proteases are hydrolytic enzymes capable of degrading proteins into small peptides and

amino acids. They account for nearly 60% of the total industrial enzyme market. Proteases

are extensively exploited commercially, in food, pharmaceutical, leather and detergent

industry.

Given their potential use there has been renewed interest in the discovery of proteases with

novel properties and a constant thrust to optimize the enzyme production. As compared to

plants and animals, microorganisms represent an attractive source of proteases as they can be

cultured in large quantities in a relatively short time by established fermentation methods, and

they produce an abundant, regular supply of the desired product. Also microbial proteases

have a longer shelf life and can be stored under less than ideal conditions for weeks without

significant loss of activity. In general microbial proteases are extracellular in nature and are

directly secreted into the fermentation broth by the producer, thus simplifying downstream

processing of the enzyme as compared to proteases obtained from plants and animals.
Materials used;

1. Tryptone soy agar.

2. Fresh culture of the bacteria isolates

3. Bacteriological agar

4. Casein

5. Conical flasks

6. Petri dishes

7. Peptone

8. Trisodium citrate

9. Na2CO3

10. MgSO4

11. K2HPO4

12. Kcl

13. NH4NO3

14. Bovine serine albumin

15. Sodium acetate

16. Calcium acetate.

17. Tween 80 solution.

Screening Procedure;

The isolates to be used ought to be screened first to be sure they still have the ability to

produce the enzyme protease. This is carried out by preparing a fresh Tryptone soy agar and

sub culturing the isolates on it and incubating at 37 degrees Celsius for 24 to 48 hours.
The colonies developed are subcultured in another prepared Tryptone soy agar which has

0.2g casein protein incorporated into it and allowed to incubate for 24 hours under same

conditions. The resulting colonies seen are noted and a mercury chloride solution is poured

on the surface of the colonies and allowed to stay for 1 minute before it is drained out. Clear

halo zones are seen around the colonies which can produce the protease enzyme. The halo

zones are measured using a metre rule to determine which has the largest diameter. It follows

that the isolate(s) with highest diameter is/are hyper producers and have the ability to produce

protease enzyme that enables it break down casein a complex protein into simpler peptides

and amino acids it can utilize. The hyper producer isolate(s) is identified on the trptone soy

agar without casein and is used for the enzyme production.

Procedure for protease production using submerged fermentation method;

1. Specified quantities of Na2CO3, MgSO4, K2HPO4, Kcl, NH4NO3, Trisodium citrate and

peptone are weighed into a 1000ml conical flask and 600ml of distilled water is added.

2. The PH of the fermentation media prepared above is adjusted to 7.3+ 0.2 ie: range (7.1-

7.5) and split into three conical flasks, 200ml each hence the three isolates we are

working with.

3. The broths in the three flasks are homogenized using a magnetic spindle bar and

magnetic hot plate stirrer and autoclaved at 121 degree Celsius and 15 Psi for 15

minutes to achieve sterility.

4. Few drops of tween 80 solution is placed into a bottle containing 150 ml distilled water

and autoclaved to ensure sterility.

5. The stock culture plates containing the hyper producer isolates are gotten, a little amount

of the sterile tween 80 is poured on the surface of the isolate colonies and a sterile

wireloop is used to carefully scrape off the colonies into the tween 80 bulk solution.

6. The tween 80 mixture containing the isolates are poured into the fermentation broths for

each isolate and swirled to mix properly.

7. The three flasks containing the fermentation broths and isolates are placed gently in a

shaker incubator of 32 degrees Celsius for 5-7 days.

8. After 7days the contents in the conical flasks are centrifuged to remove particles, the

clear solution becomes the crude enzyme solution which has to undergo a number of

purification procedures to obtain a pure enzyme.

Assay for protease enzyme activity.

A standard curve or procedure has to be established first.

 A phosphate buffer of 50 Mm, and PH 7.5 at 37 degrees Celsius is prepared by

dissolving 11.4mg/ml of KH2PO4 in distilled water

 Prepare 0.65% (w/v) BSA by dissolving 6.5mg of BSA in 1ml of the phosphate

buffer. Adjust the PH to 7.5 at 37 degrees Celsius using 0.1N NaOH or 0.1N Hcl.

 Prepare a 1:55 dilution of 110 Mm Trichloroacetic acid solution of 6.1N with

distilled water.

 Prepare 1:4 dilution of Folin.C reagent using distilled water.

 Prepare 500 Mm anhydrous Na2CO3 solution by dissolving 53mg in 1ml of

distilled water.

Using sodium acetate trihydrate and calcium acetate prepare a buffer to serve as

the enzyme diluent, adjust PH to 7.5

 To prepare the standard solutions:

 Prepare by 0.2mg in 1ml tyrosine, free base in distilled water, heat gently until

tyrosine dissolves. Cool to room temperature

 Immediately before use prepare a solution containing 0.1-0.2 units/ml of an

already known protease enzyme in the enzyme diluent prepared above

(buffer).

Tube1 Tube2 Tube3 Blank

5ml BSA 5ml BSA 5ml BSA 5ml BSA

Incubate at 37°C Incubate at 37°C for 5mins Incubate at 37°C for 5mins Incubate at

for 5mins 37°C for 5mins

1.0ml Test 0.70ml test enzyme 0.50ml Test enzyme -----no enzyme

enzyme Swirl to mix and inc at 37°C Swirl to mix and inc at 37°C Swirl to mix and

Swirl to mix and for 10 mins for 10 mins inc at 37°C for

inc at 37°C for 10 mins

10 mins.

5ml TCA 5ml TCA reagent 5ml TCA reagent 5ml TCA

reagent reagent
No enzyme 0.30ml enzyme added 0.50ml enzyme added 1.0ml enzyme

added Swirl and inc for 30 mins at Swirl and inc for 30 mins at added Swirl and

Swirl and inc for 37°C and filter 37°C and filter inc for 30 mins

30 mins at 37°C at 37°C and

and filter filter

To 4 fresh tubes;

Tube1 Tube2 Tube3 Tube4

2ml of filterate 2ml of filterate 2ml of filterate 2ml of filterate

5ml Na2CO3 5ml Na2CO3 5ml Na2CO3 5ml Na2CO3

1ml Folin C 1ml Folin C 1ml Folin C 1ml Folin C

reagent reagent reagent reagent

Swirl and incubate Swirl and incubate Swirl and incubate Swirl and

at 37°C for 30 at 37°C for 30 at 37°C for 30 incubate at 37°C

minutes. minutes. minutes. for 30 minutes.

To prepare standard curve using the standard solution prepared:

Tube1 Tube2 Tube3 Tube4 Tube5 STD Blank

0.05ml std 0.10ml std 0.20ml std 0.40ml std 0.50ml std 0.00

soln reagent soln reagent soln reagent soln reagent soln reagent

1.95ml 1.90ml 1.80ml 1.60ml 1.50ml 2.00ml

distilled distilled distilled distilled distilled distilled

water water water water water water

 5ml Na2CO3 5ml Na2CO3 5ml Na2CO3 5ml Na2CO3 5ml Na2CO3 5ml Na2CO3

1ml Folin C 1ml Folin C 1ml Folin C 1ml Folin C 1ml Folin C 1ml Folin C

reagent reagent reagent reagent reagent reagent

 Mix by swirling and incubate blanks, standards and tests at 37°C for 30 minutes.

 Filter each blank, standard and test, using a 0.45 micrometer syringe filter into

suitable cuvettes

 Record the absorbance at 660nm of each test, standard and blank solution.

For standard curve:

ΔA660nm(Standard) = ΔA660nm(Standard) - Δ660nm(Standard blank)

Plot the ΔA660nm(Standard) vs µmoles of tyrosine

For Sample determination:

ΔA660nm(Test) = ΔA660nm(Test) – ΔA660nm(Test blank)

Units/ml of enzyme = (µmoles Tyrosine equivalent released)(11)

---------------------------------------------------
(1)(10)(2)
Where;

11 = Total volume of assay in millilitres

2 = Volume (in millilitres) used in colorimetric determination

1 = Volume of enzyme used for assay

10 = Time (in minutes) of assay.

 CHAPTER FOUR

WASTE UTILIZATION AND FERMENTATION DIVISION:

Waste Utilization and Fermentation Division is one of the pioneer Divisions of the

Biotechnology Department of Federal Institute of Industrial Research, Oshodi, Nigeria.

Among the core mandates of the Division are:

1. To conduct research and development, activities that lead to the conversion of wastes

(mainly agricultural wastes) into wealth creation.

2. To carry out research and development activities that employ current fermentation

techniques to produce goods and services that will benefit Nigerians in particular and

humanity at large.

3. To cooperate with other Departments and Divisions within the Institute to disseminate

the Research and Development findings to Nigerians through

Seminars/Symposiums/Workshops

MUSHROOMS CULTIVATION

CLASSIFICATION

A mushroom (or toadstool) is the fleshy, spore-bearing fruiting body of a fungus, typically

produced above ground on soil or on its food source. The standard for the name "mushroom"

is the cultivated white button mushroom, Agaricus bisporus; hence the word "mushroom" is

most often applied to those fungi (Eumycota, Basidiomycotina, Agaricomycetes) that have a

stem (stipe), a cap (pileus), and gills (lamellae, lamella) on the underside of the cap. These gills

produce microscopic spores that help the fungus spread across the ground or its occupant

surface. "Mushroom" describes a variety of gilled fungi, with or without stems, and the term is
used even more generally, to describe both the fleshy fruiting bodies of some Ascomycotina

and the woody or leathery fruiting bodies of some Basidiomycotina, depending upon the

context of the word.

Sporocarp (fungi), Basidiocarp, and Ascocarp a polypore mushroom. Typical mushrooms are

the fruit bodies of members of the order Agaricales, whose type genus is Agaricus and type

species is the field mushroom, Agaricus campestris. However, in modern molecularly defined

classifications, not all members of the order Agaricales produce mushroom fruit bodies, and

many other gilled fungi, collectively called mushrooms, occur in other orders of the class

Agaricomycetes. For example, chanterelles are in the Cantharellales, false chianterelles such

as Gomphus are in the Gomphales, milk-cap mushrooms (Lactarius, Lactifluus) and russulas

(Russula), as well as Lentinellus, are in the Russulales, while the tough, leathery genera

Lentinus and Panus are among the Polyporales, but Neolentinus is in the Gloeophyllales, and

the little pin-mushroom genus, Rickenella, along with similar genera, are in the

Hymenochaetales. Within the main body of mushrooms, in the Agaricales, are common fungi

like the common fairy-ring mushroom, shiitake, enoki, oyster mushrooms, fly agarics and other

amanitas, magic mushrooms like species of Psilocybe, paddy husk mushrooms, shaggy manes.

Forms deviating from the standard morphology usually have more specific names, such as

bolete, puffball, stinkhorn, and morel, and gilled mushrooms themselves are often called

"agarics" in reference to their similarity to Agaricus or their order Agaricales. By extension,

the term "mushroom" can also designate the entire fungus when in culture; the thallus (called

amycelium) of species forming the fruiting bodies called mushrooms; or the species itself.

CULTIVATION OF EDIBLE/MEDICINAL MUSHROOM

Mushroom has been defined as a macro-fungus with a distinctive fruiting body which can either

be epigeous (growing on or close to the ground) or hypogeous (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.

Mushrooms are a good cash crop; they are rather easy to grow and are brimming with protein,

B vitamins and minerals, they even have medicinal properties. Time between spawning and

harvesting can be as short as three weeks. Furthermore, after the cultivation, you can still use

the substrate as a good soil conditioner and also assay and extract extra cellular enzymes.

The agricultural wastes recycling with applications in agro-food industry is one of the

biological challenging and technically demanding research in the biotechnology domain known

to mankind so far. Annually, the accumulation of huge amounts of these wastes cause serious

environmental pollution effects. Many of these lingo-cellulose wastes cause serious

environmental pollution effects. These agro-wastes are produced in significant quantities all

over the world.

To solve the environmental troubles raised by the accumulation of these organic wastes, the

most efficient way is to recycle them through biological means. As a result, the cultivation of

edible and medicinal mushrooms was applied using both the solid state cultivation and

submerged fermentation of different natural by-products of agro-food industry that provided a

fast growth as well as high biomass productivity.

These Plant wastes can be used as the main ingredients to prepare the organic composts for

edible and medicinal mushrooms growing in order to get organic food and biological active
compounds from the nutritive fungal biomass resulted after solid state cultivation or submerged

fermentation of such natural materials.

The Waste Utilization and Fermentation division section carry out research work majorly on

the cultivation of species of edible and medicinal mushroom using agricultural wastes.

MUSHROOM SPECIES CULTIVATED

 Pleurotus ostreatus

 Pleurotus pulmonarius

 Calocybe indica

MUSHROOM CULTIVATION

The major practical steps of mushroom cultivation are;

1. Selection of an acceptable mushroom species

2. Mushroom Tissue Culture

3. Preparation of Spawn

4. Preparation of Substrates to be used for cultivation

5. Spawning pasteurized substrates

6. Management of fruiting/ mushroom development

7. Harvesting and packaging of mushroom

SELECTION OF ACCEPTABLE MUSHROOM SPECIES

Before any decision to cultivate a particular mushroom is made, it is important to determine if

that species possess organoleptic qualities acceptable to the local population. The most

common method used, is the “Ask Around” or you carry out Molecular characterization.

MUSHROOM TISSUE CULTURE

After selecting your Mushroom species of interest, you progress to your Mushroom Tissue

culture which is done on a culture media. A Culture medium is a solid or liquid that supports

the growth of microorganisms or cells.

 PDA (Potato dextrose Agar) – it is the simplest and the most popular medium for

growing mycelia of most cultivated mushrooms. You prepare your PDA according to

your manufacturer’s instruction.

STEPS FOR TISSUE CULTURE

 A freshly harvested Mushroom is gotten and cleaned with 70% ethanol.

 The mushroom is then split into half by hand longitudinally under aseptic conditions.

 Some inside tissue is taken with a loop and placed on your Potato Dextrose Agar plate

which is your medium.

 As soon as the tissue is transferred, the plate is closed and incubated in the incubator

between 2 OC – 34OC depending on the mushroom species

 Some white, delicate Mycelia will be produced from the small piece of tissue within

two to three days.

  After about ten days the mycelium will grow rapidly and cover the surface of the agar

medium.

Then it is ready to transfer to the spawn substrate for the production of spawn.

PREPARATION OF SPAWN

What spawn is to mushroom is what seed is to plant. Mushroom Spawn is a medium through

which the mycelium of a fruiting culture has grown and which serves as the inoculum of ‘seed’

for the substrate in mushroom cultivation. Mushroom spawn preparation is one of the critical

points in mushroom cultivation as it could lead to failure to achieve a satisfactory harvest, or

no yield at all. Basically, spawn production is nothing more than putting mycelium of the

desired mushroom in suitable sterilised substrates under aseptic conditions.

SPAWN SUBSTRATES

A number of materials, mostly agricultural wastes, can be used to prepare mushroom spawn.

Some of the materials are Cereal grains (Sorghum or wheat), cotton wastes, sawdust, they can

all be used as spawn substrate.

PREPARATION OF MOTHER SPAWN – using Sorghum grains.

 The Sorghum grains are washed to remove the stones and debris

 After washing, it is boiled in water for about 10 to 15 minutes until they expand but not

quite broken. (CaCo3 is added to the boiling water so as to serve as a buffer and regulate

the pH level and also to prevent caking of grains after sterilization)

 The grains are drained and allowed to cool. CaSO4 is added to speed up the drying

process.

 Grains are then loosely packed in bottles which are 2/3 full and covered with Aluminum

foil and tightened with rubber band.

  The bottled grains are sterilized in an autoclave for 1 hour 30 minutes at 121°C and

then cooled prior to inoculation.

 Inoculation is done in an aseptic environment in an aseptic way preferably in a room

without air movement (close doors and windows).

 The pure culture spawn from your tissue culture is inoculated from your plates into your

bottles containing your sorghum grains aseptically and then closed with the aluminium

foil and tied with a rubber band.

 After inoculation, the bottles containing the inoculum (Mushroom tissue) are then

placed in a dark cupboard under room temperature for ramification.

Note: Autoclaved substrates can only be justified for spawn production if it is properly done.

Otherwise there will be wastage of energy and money through contamination losses.

Spawn production.

PREPARATION OF SUBSTRATE FOR MUSHROOM CULTIVATION.

Mushroom substrate can be defined as a material which supports the growth, development, and

fruiting of mushroom mycelium. Many materials regarded as wastes can be used for mushroom
cultivation. Materials like Sawdust, Rice bran, Wheat bran, wood shavings, Cotton wastes, Dry

banana leaves amongst others. Some of these wastes could be combined or used individually.

PREPARATION OF SUBSTRATES Using Sawdust + Rice Bran + banana leaves for

example.

 Your dry banana leaves are shredded and soaked in water containing CaCo3 for 24

hours.

 Sawdust and rice bran are mixed in a 4:1 ratio, then you add your banana leaves and

mix

 Add CaCO3(gypsum) to the mixture so as to minimize the greasiness compost normally

tend to have and also water so as to increase the moisture level but not make it

waterlogged

 Once the pile is wetted and formed, aerobic fermentation(composting) commences as

a result of growth and reproduction of microorganisms which naturally occur in bulk

ingredients.

 Turning and watering are done at approximately 2-3days intervals. Water addition is

critical since too much water will exclude oxygen by occupying the pore space, and too

little can limit the growth of bacteria and fungi.

 After composting, the next phase is pasteurization, which is necessary to kill any

insects, nematodes, pest fungi, or other pests that may be present in the compost

 Bag your compost and pasteurize in a local drum between 3 – 5 hours prior to

inoculation.
BULK PASTEURISATION BY STEAM

This method kills the unwanted organisms but keeps the favourable ones alive. To achieve this,

a temperature of 60 ºC to 70 ºC has to be sustained for at least 5 hours; after which most pests

and diseases (contaminants) will be eliminated.

Materials and equipment required:

 Substrate material

 Substrate containers (e.g. plastic bags)

 Oil drum and burner

SPAWNING PASTEURIZED SUBSTRATES/SPAWN RUN

Following pasteurization by steam to kill off potential competitive microorganisms,

Inoculation of ramified spawns is done aseptically into the pasteurized bags. Once the bags are

spawned they are placed on shelves in the incubation rooms. Depending on the strain and

temperature, the mycelium will colonise the substrate in two or three weeks, this is the phase

which the mycelium grows from the spawn and permeates into the substrate.

The mycelium will colonize the substrate and use the available nutrients, this is commonly

referred to as the spawn run. During spawn run stage the mycelium will grow through the

substrate, the spawn run time is different for each species and depends on the size of the bag,

amount of spawn used, the strain used and the temperature.

After the mycelium totally colonizes the substrate, next you move to the fruiting room where

you expose by creating little holes and also for watering the bags for fruiting.
MANAGEMENT OF FRUITING/MUSHROOM DEVELOPMENT

The ramified substrates are then taken to the fruiting room for exposure, under suitable

environmental conditions such as temperature, ventilation, light and humidity. Primordial

formation occurs and then followed by the production of fruiting bodies, Good control

humidity is very important for all types of mushroom. Humidity is kept high by constantly

spraying water, however no water should be sprayed directly onto mushrooms that are ready

for picking. Their shelf life will decrease drastically if they become too wet

HARVESTING MUSHROOMS/PACKAGING

Harvesting is carried out at different maturation stages depending on the mushroom species

and upon consumer preferences and market value. The mushroom fruiting body is removed

carefully from the bag. The mushrooms are ready for harvesting in five days. Harvesting is

performed by gently pulling or twisting the mushrooms from the substrate. Only very little

substrate should be pulled out.

Freshly harvested mushrooms must be kept refrigerated. To prolong the shelf life of

mushrooms, it is important that mushrooms “breathe” after harvest, so storage in a non-waxed

paper bag or perforated plastic bag is advisable production.

Spawn substrates. Sprouting mushrooms from substrates.

ECONOMIC IMPORTANCE OF MUSHROOM CULTIVATED

Nutritional value: mushroom is a source of energy food, carbohydrate, protein and fats.

Mushroom rank below animal meats but above most other foods including milk which is

animal product. Mushroom protein contains all the nine essential amino acids required for man

including fats, phosphorus, iron and vitamins like thiamine, riboflavin, ascorbic acid,

ergosterine and niacin, they are low in calories, carbohydrate and calcium. Mushroom also

contains high proportion of unsaturated fat. E.g. Auriculariasp, Agaricussp, Morchela

esculenta

Biotechnology: Mushroom enhances human health. It is concerned with mushroom product s

and encompasses the principle of fermentation technology, mushroom biology and bioprocess.

Mushroom products have a generalized or ionic effect which in turns may act prophylactically

by increasing resistance to disease in human, for balancing of nutrients in diet, and enhances

immune system. E.g. Agaricusbisporus, Lentinulasp, Pleuropus sp.Torulopsisutilis,

Saccharomyces cerevisiae
Bioremediation: turning waste into wealth, this area deals mainly with the aspects of benefits

to the earth from the activities of mushroom mycelium. Environmental contamination can be

ameliorated by the application of mushroom mycelia technology.

 Use of bioconversion process to transform the polluting substances into valuable food

stuffs, i.e. proper treatment and reutilization of spent substrates/compost to eliminate

pollution problems. E.g. Oyster mushroom

 Use of mushroom mycelia as tools for healing soil called “Mycoremediation”,

mycofiltration (filter water), mycoforestry in ecoforestry policy.

Medicinal properties: The mycelium of the fruiting body of mushroom serves as nutriceutical

for disease treatment and antibiotics, tonics, anticancers.

E.g. Ganoderma applanatum for antibacterial and anti-inflamatory, antitumor, Lentinula

edodes for anti candida, Pleurotus ostreatus for antiviral and promotes health.
FERMENTATION OF MOLASSES GOTTEN FROM BREWERIES FOR ETHANOL

PRODUCTION.

The fermentation process was performed by the yeast Saccharomyces cerevisiae.

Preparation of the substrate:

Preparation of molasses consisted of its dilution with distilled water in a ratio of 1:5 in

a beaker. The diluted molasses were then clarified by acidifying it to a PH of 4.5 with

concentrated sulphuric acid followed by boiling on a hot plate stirrer for about 30 minutes with

constant stirring by the magnetic spindle bar stirrer. This was left to cool, and poured into a

measuring cylinder and covered. It was left overnight to settle some of the impurities such as

peptides and proteins. The supernatant was decanted after settling and diluted with water to

20% sugar content, then salts, ammonium sulphate and ammonium hydrogen phosphate were

added in the amount of 0.85% and 0.12% (w/v) respectively. The PH was again adjusted to 4.5

– 5.0, then sterilized at 150°C for 30 minutes and cooled to 27°C. The medium was now ready

for inoculation with the yeast for fermentation.

The resulting solution is received in a large tank and yeast is added to it at 30°C and

kept for 2 to 3 days. During this period, enzymes sucrase and zymase which are present in

yeast, convert sugar into ethyl alcohol. As days go by, the sugar content of the molasses

decreases giving rise subsequently to alcohol increase

C12H22O11+H2O C6H12O6+C6H12O6

C6H12O6 C2H5OH+2CO2

Ethanol obtained by the fermentation is called WASH, which is about 15% - 18% pure.

By using fractional distillation technique, it is further purified to about 92% pure alcohol which

is known as rectified spirit or commercial alcohol.

FOOD SAFETY AND QUALITY MANAGEMENT SYSTEM DIVISION:

This is a pioneer arm of the biotechnology unit This department handles chemical,

microbiological and toxicological analysis of raw materials, semi-finished products as well as

finished products. Analysis are run daily of food samples to check if regulatory, statutory and

food safety standard requirements are met.

A number of tests are carried out on food materials and portable water and these test depend

on certain critera:

 History of the sample (type of raw materials used)

 The type of processes used for the production

 Nature of the food material

Packaged food material have to undergo coliform test to check for contaminations from

improper handling by factory workers.

Water samples are tested for pathogens like Salmonella, Shigella, coliforms etc.

Foods produced in vacuums like canned and tinned milk, fish etc are analysed for clostridium,

bacillus (spore formers)

Regulatory agencies have established a food safety limit which is reviewed often, this guide

gives the total aerobic count of bacteria and yeast/mold allowed in food samples. Exceeding

these limits implies an unsafe food material and as such a defect in good manufacturing

practices.
Problems Encountered;

The students industrial work experience scheme was a wonderful experience for me, one that

has greatly added value to my life.

First, my biggest challenge was the distance i was coming from. I had to take two to three drops

every morning before I get to work and this is chiefly because i didn’t get placement in any

organization close by either because their facilities are insufficient or they don’t accept students

at all.

Recommendation:

SIWES managers should partner with companies so that trainees can be accepted into their

institution for practical training and reduce the stress on students seeking a place of attachment

(even without allowances and benefits) which most times end up fruitless.

The SIWES and ITF management should endeavour to reach as many companies that have

students attached to them for training to see how they are being trained, their welfare and how

the program is being monitored by supervisors.

The management should try and make it a condition for companies to pay IT students, as most

times the students are used for errands within and outside the premises.

The management should organize orientation programs for students prior to their attachment

to further enlighten them on the relevance of hands on practical knowledge in their field.