STUDY ON THE EFFECT OF MOISTURE VARIATIONS ON

THE MICROBIOLOGICAL QUALITY AND SAFETY OF

MALTED SOY FORTIFIED GARRI

BY
NWAUDO GIDEON NNAYERE
20101711623

DEPARTMENT OF FOOD SCIENCE & TECHNOLOGY, SCHOOL

OF ENGINEERING & ENGINEERING TECHNOLOGY
FEDERAL UNIVERSITY OF TECHNOLOGY, OWERRI.

OCTOBER, 2016

CERTIFICATION
This is to certify that this research work titled THE EFFECT OF
MOISTURE

VARIATIONS

ON

THE

MICROBIOLOGICAL

QUALITY AND SAFETY OF MALTED SOY FORTIFIED GARRI

carried out by NWAUDO, GIDEON N with registration number
20101711623

under the supervision of Dr (Mrs) N.N Ahaotu of the

department of Food Science and Technology in partial fulfillment of the

requirement for the award of Bachelor of Technology (B.TECH) degree in
Food Science and Technology.
-------------------------------

-------------------------

Dr. (Mrs) N. N. Ahaotu

DATE

Supervisor

--------------------------------

-------------------------

DR. M.C Osuji.

DATE

(Head of Department)

--------------------------------

----------------------------

External Examinar

DATE

DEDICATION
This work is dedicated to GOD almighty for his superabundant provision to
this stage of my academic pursuit and also to my parents and siblings for
their endless Love, prayer and support financially and otherwise.

ACKNOWLEDGMENT

First and foremost, I want to thank God Almighty for his super abundant
and immeasurable love and favours during the course of my study in
Federal Unoversity of Technology Owerri.
I thank specially my efficient and respectable supervisor, Dr. (Mrs) N.N
Ahaotu, for her understanding, patients, constructive criticism which
contributed greatly to the success of this work and words of encouragements
throughout my stay in this school.
My appreciation also goes to my Head of Department for his understanding
and open mindedness. My sincere thanks goes to my class adviser who
doubles as my supervisor, Dr. (Mrs.) N.N. Ahaotu, for her encouragements
and motherly response during this research and also to all my lecturers for
impacting knowledge and also for their support.
Im greatly indebted to my loving parents Mr. and Mrs. Duru Nwaudo for
their numerous supports both financially, spiritually and morally. I
profoundly

appreciate

also

my

uncle

Chigozie

Ogbuji,

for

his

encouragements and moral supports.

Im not forgetting my lovely siblings, Mrs Duru Grace, Glory and
Emmanuel Nwaudo I say a big thank you for all your supports and prayers.
And to the laboratory of Food Science and Technology Owerri, for their
laboratory assistance. I also thank the farm manager, Mr. Vincent of

National Root Crop Research Institute Umudike and Mrs Chikere for
providing the raw material. And Mr Justice for his laboratory assistance.
Finally I want to use this medium to acknowledge my project partner
Umejesi Joseph and my friends, Opara Assumpta, Nworie Chinazom,
Nzeribe Ikechukwu, Emmanuel Anthony, Iwunze Chuka and others for their
encouragement.

TABLE OF CONTENTS

COVER PAGE................................................................................................i
CERTIFICATION..........................................................................................ii
DEDICATION...............................................................................................iii
ACKNOWLEDGMENTS.............................................................................iv
TABLE OF CONTENTS..............................................................................vi
LIST OF TABLES.......................................................................................viii
LIST OF FIGURES.......................................................................................ix
ABSTRACT...................................................................................................x
CHAPTER ONE
INTRODUCTION..........................................................................................1
1.1
1.2

BACKGROUND OF STUDY..............................................................1
PROBLEM STATEMENT SAFETY OF MALTED SOY

FORTIFIED GARRI AT DIFFERENT MOISTURE.....................................3

1.3 JUSTIFICATION OF THE STUDY........................................................4
1.4

OBJECTIVES OF THE STUDY..........................................................4

CHAPTER TWO
LITERATURE REVIEW...............................................................................5
6

2.1

HISTORICAL SOURCE AND DEVELOPMENT OF CASSAVA......5

2.2

UTILIZATION OF CASSAVA AND ITS PRODUCTS.......................8

2.3 STORAGE OF CASSAVA AND CASSAVA PRODUCT......................11

2.4 PROCESSING OF CASSAVA ROOT TUBERS...................................12
2.5

THE EFFECT OF PROCESSING ON THE CYANIDE CONTENT

OF CASSAVA..............................................................................................13
2.6 PROCESSING OF CASSAVA TUBERS...............................................14

CHAPTER THREE
MATERIALS AND METHOD....................................................................21
3.1 MATERIALS..........................................................................................21
3.2

PREPARATION OF MALTED SOYBEAN EXTRACT FORTIFIED

(FORTIFIED GARRI) GARRI....................................................................21

3.3

SAMPLE PREPARATION FOR GARI (UNFORTIFIED I.E

WHITE)........................................................................................................22
3.4

MOISTURE VARIATION..................................................................23

CHAPTER FOUR
RESULTS AND DISCUSSION...................................................................29
CHAPTER FIVE
7

DISCUSSION...............................................................................................34
5.1

MICROBIAL COUNT ON CASSAVA GARRI..............................34

5.2

CONCLUSION...................................................................................36

5.3

RECOMENDATION..........................................................................37

REFERENCES.............................................................................................38
APPENDIX 1...............................................................................................42

LIST OF TABLES

Table
4.1

Page
Frequency of Fungi (Mould/Yeast) species isolated
from garri samples after ten weeks of storage.-

4.2

4.4

33

Garri produced from Manihot esculenta (TME 419)

34

Mean value of the total Bacteria count from

Mean value of the total Mould and Yeast count

from Garri produced from Manihot esculenta (TME 419)

4.5

32

Frequency of Bacterial species isolated from garri samples after

ten weeks of storage

4.3

35

Mean value of the total Fungi count from Garri

produced from Manihot esculenta (TME 419)

36

LIST OF FIGURES
Figure

Page

3.1

The flow diagram for the preparation of gari -

3.2

Flow diagram of production of malted soy fortified gari -

3.3

Flow diagram for the preparation of soybean flour

4.1:

Effect of Moisture Variation on the Total Bacteria

Count on the gari samples

4.2:

24
25

26

29

30

31

Effect of moisture variation on the yeast/mould

growth on the gari samples

4.3:

Effect of moisture variation on the total fungi count

on the gari samples

10

ABSTRACT
This study evaluated the effect of moisture variation on the microbiological
quality of stored malted soy fortified garri. Soy fortified garri produced
from the cassava cultivar TME 419 having different moisture levels (8%,
10%, and 12%) was stored for ten (10) weeks. The samples were analysed
to ascertain its microbiological quality within the storage period. There was
a progressive increase in the total plate count for mesophillic bacteria
throughout the storage period from 0.3x 103cfu/ml in week zero to 53 x
103cfu/ml in week 10. The mean values of the microbial load of the fortified
gari stored at different moisture levels were higher than the mean value of
the control samples. There was a significant variation in the mean values
obtained for yeast/mould counts for the fortified samples and the control
samples. The various data from the different samples showed a significant
increase in microbial load with time and moisture content of the stored
samples, although the microbial load values are still within acceptable limits
for the different moisture levels studied. Both the fortified samples and the
control samples shared the same bacterial specie isolates which includes;
Staphylococcus spp, Pseudomonas spp, Micrococcus spp, Acinetobacter spp
and Achromobacter spp. after 10 weeks storage except for the incidence of
Bacillus spp isolated from the fortified gari sample and not present in the
control sample. The fungal species associated with the fortified sample at
the end of the ten weeks storage included; Rhizopus spp, Cladosporum spp,
and Rhodotorula spp while the control sample had Aspergillus spp,
Penicillium spp, Cladosporum spp and Rhodotorula spp.
Keywords: Moisture variation, Microbial count, fortified, gari

11

CHAPTER ONE

1.1

INTRODUCTION
BACKGROUND OF STUDY

Cassava (Manihot esculenta crantz) belongs to the genus manihot of the

natural order euphoriacea (Onyenuga, 1998) thus is large family which
yields latex (Ngoka, 1977) Lancaster et al., (1982) reported that cassava is a
perennial shrub ranging in height from 1.5,with branching stem, green, pale
or dark grey or brown coloured. cassava a root are generally from15-100cm
long and 3-15cmwide, the are conical, cylindrical or oval, with a pink,
cream or coffee coloured peel that is covered by a thin brown bark (Macrea
et al., 1993) cassava is a woody plant, which endures drought and grows
well in areas of medium rainfall (Ngoka,1997). The crop gives its optimum
yield when grown on well drained loam or sandy soil with plenty of organic
matter.
Cassava can also be known as manio or tapico root, yucca (Spanish)
mandioca (Portuguese), arrow root (Brazilian) rogo (Hausa), akpu (Ibo),
gbaguda or ege (Yoruba), (Onyenuga, 1968).
Cassava is often classified by the amount of bitter substance in the root and
bitter and sweet can be found. The bitterness is caused by cyanogenic
glucoside contained root. Cyanide is released upon crushing the roots as
would in chewing. (habor C.I and ogundu, E.C 2009) The presence of
12

cyanide in the roots is a natural form of protection for the plant soil and
climatic conditions determine the amount of this compound in the roots.
In general bitter cassava has high cyanide content of 100mg/kg HCN
while sweet cassava has a lower value of les than 100mg/kg HCN.
Among the tropical root and tuber crop grown worldwide, cassava ranks
first in per capital consumption, it provides 30% of the total calorie intake
(Scott et al, 1991).
Cassava has low protein content and for cassava product to be nutritionally
adequate it must be fortified or supplemented. (Edem D.O Ayatse and ham
E.H 2001) With a protein rich in food such as soy flour (Almazan, 1987).
When eaten as a staple food as a staple food, cassava as normally served
with a protein rich dish such as traditional soup with meat or fish and leafy
vegetable (Ihekoronye and Ngoddy, 1985).
Cassava tubers are extremely perishable and cannot be stored for more than
a few days. The deterioration of cassava is caused by microbial infection
such as minor wet rot and physiological factors such as moisture loss
(Kwattia, 1986) both root and leaf contains varying amounts of cyanide
which is toxic to human and animals. Cassava is an important major food
crop in Nigeria estimated to supply about 70% of daily calories of over
50millions people in Nigeria
13

1.2

PROBLEM STATEMENT

Widespread malnutrition with ever increasing protein gap in our country has
necessitated the research for alternative protein. That brought about the
additional attachment of protein supplement which is soy bean flour, It has
been estimated in this study to investigate various moisture storage and
effective of Ziploc packaging material to withstand environmental
organisms and the best storage duration for processed cassava tubers since
its highly effected by fungi, bacteria, mould (oyeniran. JO (1995) and other
organism which tend to reduce the shelf stability which could bring about
mass spoilage and health risk to consumers

1.3 JUSTIFICATION OF THE STUDY

Information obtained from this research will help in improving the shelf life
of this very nutritious food due to widespread malnutrition with ever
increasing protein gap in our country this product will help to bridge the
protein and overall nutrients gap this research work would also enlighten
people on the functional microbiological properties of malted soy fortified
garri

1.4

OBJECTIVES OF THE STUDY

14

1 To determine the effect of different moisture levels on the microbiological

quality of malted soy fortified gari.
2 To determine the safety of the product after ten weeks of storage
3 To isolate the implicating spoilage organisms

15

CHAPTER TWO

LITERATURE REVIEW
2.1 HISTORICAL SOURCE AND DEVELOPMENT OF
CASSAVA
Cassava originated from central and South American and spread rapidly
arriving on the west coast of African via gulf of Benin and River Congo at
The end of the sixteenth century the cop spread in the east coast via. The
Reunion Island inadequate and Zanzibar at the end of the eighteenth
century. By the early 1800s cassava arrived in India the land Indonesia
(Macrae et al., 1993).
Cassava is known as one of the major staple food crop. Many technical
factors have earned this reputation for cassava. Cassava is an extremely
undemanding crop which is able to grow under a variety of climatic and soil
condition (NRCRI, 1995). Cassava is tolerant to drought and once
established, cassava unlike other crop has no critical period when lack rain
will cause crop failure (Kwatia, 1986).
Cassava grows in all types of soil but prefers a sandy or sandy loam soil
with hard pan (Impenetrable layer) about 30- 40cm deep tare desirable
because they prevent deep penetration of roots which aids harvest. Land
preparation is carried out after a good factors, field cultivation is done in
ridges 90- 100 cm apart or in heaps or mounds. planting may be carried out
16

at anytime of the year provided there is sufficient moisture to aid sprouting

(IITA,1990) propagation is by woody steam pieces called stakes, the
optimum daily temperature for cassava is between

18-35 0c and the

minimum temperature it can tolerate is 100c even a light frost kill the crop.
Cassava is drought resistant, during this period the plant drops its leaves,
which helps prevents rapid water loss from the roots, the leaves quickly
grow once rainfall occurs soil with good agronomic practices, up to 50per
hectare can be produced in 12 months, as a worldwide basis, the crop more
frequently intercropped will cassava, maize, cowpea sorghum and millets
(Macrae et al, 1993).
Cassava root generally from 15-100cm long and 3-15cm wide. They are
cylindrical, conical or oval, with a coffee, pink, or cream colored peel,
which is covered by brown bark. The parenchyma, the edible portion of the
fresh root comprise appropriately 85% and is generally white, cream or
yellow colour (Macrae et. al 1993).
2.1.1

Storage of Cassava Products

The cassava root has no fixed period of optimum maturity. The woody plant
is perennial and starch deposition will continue for many years. In most
ecosystem (Macrea et al., 1993). Some root mature within 12-18 months,
others within 6-7 months, tubers may be left in the ground for up to 2 years.
After that the nutrient values of the tubers will decline and the tissue will
17

become fibrous (IITA, 1990) pest and disease of cassava are grasshoppers,
Zonocous vangatus rodent, wild pigs white fly, Benisa specie (IITA 1990).
Freshly harvested cassava roots have the shortest post harvest life of any of
the major staple food crops. Roots become inedible within 24-72 hours after
harvest due to rapid physiological (0giehor I.S Ikenebomeh Mj (2005)
deterioration. This deterioration is a major constrain for Industrial
processing of fresh roots and for marketing them to distant urban centers
(Macrae et al, 1993).
Ihekoronye and Ngoddy (1985) reported that the appropriate composition of
the cassava tube is:
Starch (20-30%), Protein (2-3%), water (75- 80%), fat (0.1%), fiber (1.0%),
ash (1-1.5%), vitamin c (35mg/100 fresh weight) calcium (33mg/100g
edible portion niacin (trace), thiamine (trace)
The cassava tubers consist mainly of carbohydrate to about 90% as dry wet
basis (Key, 1973) protein content is at almost 3% and low in methionine.
Generally, cassava plant is a highly efficient producer of carbohydrate,
mainly in starch flour. It is fourth most important source of calories in the
human diet in tropical region of the world where it is consume in a wide
variety of forms. (Olaniyan (1991)

18

2.2

UTILIZATION OF CASSAVA AND ITS PRODUCTS

Many food preparation in African utilizes cassava tuber, it a good sources of

calories in a meal. In African, processed cassava in the form of flour, wet
pulp for drinking or garri can be cooked and eaten in three form base on the
different ethnic group in Nigeria fufu, eba and chikwargue
2.2.1 Garri
Garri can be eaten in different ways; cold water can be added to it and
mixed as a meal. It can be mixed with hot water to form a sticky mass (eba)
o, which is eaten with soup or stew. It can be moisturized and mixed with an
oily stew containing onion, tomatoes and spices. This mixture is highly
fixed and is known as garri forflor, garri is steamed to prepare food called
attieke in the ivory coast and Togo. Eba is a very popular form of cassava
food in Nigeria and is gaining popularly in Cameroon, Benin, Ghana and
Liberia because of it easy reconstitution into convenient food. These
qualities will be of even greater importance as urbanization advances and
life becomes busier. In African (Lancaster et al, 1988, Hahn, 1989)
2.2.2 Fufu
Fufu is a kind of porridge, strong paste or sticky dough made from cassava
flour derived either directly from sun dried root pieces or from fermented
and dried pulp. Fufu is the most common product consumed in African. The

19

fufu groups include amala. In Nigeria, tolo in Guinea, fufu in Zaire, the
Cameroon, congo Ugali and Kowon or atap in Uganda and Tanzania,
nchima In Mozambique, nsima In Malawi, Ubugali in Rwanda (Hahn,
1989).
2.2.3 Chikwangue
Chikwangue: Is very stiff paste and it is much stiffer than fufu and eba, the
shape size and texture of the chickwangue food group vary among
countries. Bobolo and myondos in Cameroon are prepared in essentially the
same way as chiekwanque, although shapes and sizes are different and thus
belongs to the Chikwangue food group (Hahn, 1989).
2.2.4 Livestock Feed
There is a considerable potential for using cassava feed nations in local
livestock industries. Since 1960s some countries have used chips in
compounds animals feed because of the high energy content and low prices
of cassava (IITA, 1990). The various forms it could be used as are suggested
by Akonoda and Arene, (1989) is discussed below.
In Brazil, whole (root, stem, and leaves) are ground and dried and fed to
ruminants. Leaves have a high protein content(20-30%) but should be
grown for fodder only on a small farms where animals dung is returned to
the soil as manure to avoid soil depletion(large scale).
20

Boiled roots, used to fatten pig, 50% of feed 17-35kg weight and up to 70%
for heavier pigs (small to large scale).

2.2.5 Cassava Root Meal

Cassava root meal, ground, fermented cassava roots (including the peel) are
used at the NRCRI as a substitute for maize in poultry feed on a ratio of 70100% for broilers and 50% for layers. A methionine supplement is essential
to ensure excellent result and fertility used in the food industry in many
preparations,

including

sauces,

gravies,

mustard

powders

glucose

production. Confectionary and baking products it is also used as jelly or

thickening agent. It is used extensively in the manufacture of adhesives
dextrin, paste and as a filter in the manufacturing of paints, in the textile
industry; it is used for wrap sizing cloth and the finishing (IITA, 1990).

2.3 STORAGE OF CASSAVA AND CASSAVA PRODUCT

Cassava tubers are extremely perishable, they can be kept underground prior
to harvest for up to about 2years but once they have been harvested they
begin to deteriorate within 40-48hrs. The deterioration is caused by the
physiological changes and subsequently by rot and decay (Achinewhu,
1991).
21

Deterioration of cassava has an adverse effect on the processed product and

thus the crop must be stored properly. In an environment safe and free of
micro organism or with a packaging material that can prevent micro
organism to a large extent.

2.4 PROCESSING OF CASSAVA ROOT TUBERS

Fresh cassava root cannot be stored for long because the rot within 3-4days
after harvest. Being bulky with about 70% moisture content, transportation
of the root from rural to urban areas for marketing is very difficult and
expensive. Both root and leaf contain varying amount of cyanide, which is
toxic to human and animal. Thus there is a need to process cassava into
various products for these reasons (Hahn, 1989).
1) To reduce the content of cyanide
2) To diversify the food and other uses of cassava leading to market
expansion.
3) To reduce bulk and hence lower transportation cost.
4) To reduce a more stable product, capable of being stored in tropical
environment for extended period with no decrease.
5) To provide food and raw material for small and medium scale cassava
based rural industries (Macrae et al, 1993).
22

2.5

THE EFFECT OF PROCESSING ON THE CYANIDE

CONTENT OF CASSAVA

Cassava is a staple food in Africa and has a potential livestock feed. A

constraint to its use is due to the presence of cyanide in the whole plant. A
cultivar is considered innocuous or safe if the total cyanide content in
HCNmg/kg fresh weight tuber is less than 50.moderately poisonous if 50100 and dangerously poisonous if it contains more than 100.
Processing of cassava tuber and leaves reduces cyanide concentration
therefore its toxicity variation in the preparation exist from time to time and
place to place due to subtle difference is preferred, taste, texture, appearance
and digestibility (Almazan, 1987).

2.5.1

Effects of Fermentation during Cassava Processing

Fermentation may be either aerobic or anaerobic. Tissue disintegration in

the presence of excess moisture during grating or fermenting in water
permits the rapid hydrolysis of glycosides effectively reduces both free and
residual in the product (Hahn and keyser, 1985; Almazan, 1987). When garri
is converted to eba, its HCN is further reduced to even safer level. However
Hahn (1985) reported that cyanide reduction of 100% was achieved by
processing roots into chikwangue.

23

2.5.2 Effects of Drying

Prior to milling of flour, peeled tuber are cut into various sizes and shapes
then either directly sun dried or fermented for a few days before sun drying,
generally sun drying is more effective than oven drying in removing cyanide
(Almazan, 1987), Hahn (1985) reported that peeled cut pieces of root gave
HCN concentration lower than 10mg/kg.

2.6 PROCESSING OF CASSAVA TUBERS

Cassava is processed into many different parts which several major ones are
described below including how they are processed.
2.6.1 Garri
Garri can be defined as a product gotten from cassava through a process of
fermentation and toasting its been consumed in several means by different
ethnics groups
The cassava roots are hand peeled then grounded into a pulp often, using a
roughly perforated iron sheet. This pulp or mash is placed on Hessian sacks
under heavy weight (logs, rocks) to squeeze out excess water. The mash is
left to drain for 1-6 days during which natural fermentation occurs. The
pressed cake is shredded by hand found sieve to remove fiber and large
lumps. This sieve mash is toasted (garified) in day or iron vessels over a
wood fire until the starch gelatinizes and the moisture content is reduced to
24

15%. The mash must be stirred constantly to avoid sticking or banning.

Palm oil may be added to facilitate this operation and imparts a yellow
colour to the final product (Macrae et al, 1993).
2.6.2

Farina De Manioc

To prepare it, freshly-dug roots are first washed and peeled the reduced to a
pulpy mass using a greater made of a float piece of wood studded with sharp
stones. The mass is them put into a cylindrical basketry press, the the
tipiti. After a few hours in the press, the pulp is removed and forced
through the sieve, then put into a shallow basin over a flow fire and stirred
until it is toasted (Ihekoronye and Ngoddy, 1985).
2.6.3 Shelf Stability Of Cassava Products
Cassava and root tubers related product has very high yield of moisture in it
in such that spoilage organism find them inviting, this water activity tends to
reduce the shelf life if its not control to a large extent by reduction of the
moisture content which could be by the use of heat treatment or radiation to
bring it to a dry matter state in as to reduce production loses in extension of
the shelf life
2.6.4 Soybean
Soybean

and soy foods contains a variety of bioactive component,

including saponins, protease inhibitors, phytic acid, and iso-flavones

25

belongs to a class of compound generally known as phytoestrongens, plant

compound that have estrogen like structures.
The original interest of soy was fueled by geographic epidemiology the
observation that population that consume a lot of soy, particularly those in
the eastern Asia have less breast cancer, prostate cancer, and cardiovascular
disease, and fewer bone fractures. Additionally, women in the population
report fewer menopausal symptoms, such as hot flashes, and both men and
women have a lower incidence of aging related brain disease development,
and diets is a major lifestyle factor, traditional Asian diets drew considerable
attention.
2.6.5 Soybean Nutritional Properties
1. Soybeans are good sources of protein, lipid and other nutrients Soy
protein products can be substitute for animal Protein because, unlike
some other beans, soy offers a complete protein profile utilization.
Soybean contain all the essential amino acids (except methionine)
which must be supplied in the diet because they cannot be
synthesized by the human body. Soy protein products can replace
animal based foods which also have complete protein but tend to
contain more fat, especially saturated fat without requiring major
adjustments elsewhere in the diet. Proteins and lipids, some vitamins
and minerals, are major nutritionally important compounds of
26

soybeans although carbohydrates are major constituents quantitatively

Soybean are consumed mainly because of its protein properties which
is about 38-44% larger than the protein content of other legumes, and
20-30% and much larger than that of cereals, 8-15% This large
amount of protein in soy beans along with the high biological value
(BV) increases their value as feedstuff and is one reason given it
economic advantage over other seeds.
2.6.6

Soybean Minerals

Dry Soybean has an ash content of about 5%, which is quite considerable.
The major forms of

minerals in Soybean are sulphates, phosphates, and

carbonates. Potassium is found in the Soybean in the highest concentration,

followed by phosphorus, magnesium, sulphur, iron, zinc, manganese,
copper, molybdenum, fluoride, chromium, selenium, cobalts, cadmium,
lead and more they range 001-140 ppm. Like other components, minerals in
Soybean.
2.6.7

Microbiological Profile

Microbiological analysis of food is the determination of the major

organisms acting on the food with respect microbial counts, identification of
organisms which includes bacteria, fungi (mold and yeast). These organisms

27

important in foods for their ability to cause food-borne diseases, food

spoilage and to produce food and food ingredients (obatolu and osho, 1992).
2.6.8 Microbial Counts
Microbial counts in food are the estimation of living and dead cells. In food
or living cells only, total cell counts and total viable counts aids the
determination of microbial counts in food.
Total cells counts gives an estimate of actual cells present in a sample and
not the numbers of colony forming units (assuming each viable cell
produces a colony) as viable count, Thus the total viable counts depend on
the ability of the organisms to divide on the solid matter and form the viable
colonies, This shows that viable counts solely estimate the number of
colony forming units without dead cells.
In general, counting in microbiological analysis refers to determining the
amounts of colony forming units (CFU) or viable microbial cells present in
a unit volume or weight of sample more so, enumerating the number of
colonies in agar places may also be referred to as counting (Oyewole, 1991).
2.6.9 Identification of Micro-Organisms
This amount to the characterization of micro-organism based on their
cultural (colonial features) morphological (cell features), biochemical
features or characteristics molecular/genetic features Identification normally
28

refers to naming the micro-0rganism genus and preferably the species as

well culturing techniques /culture characteristics and the identification of
micro organisms culture specifically includes colony appearance, colony
forms (shape), colony elevation, colony margin, optical density, colour
(pigmentation), colour, odour, colony consistency etc.

29

CHAPTER THREE
MATERIALS AND METHOD
3.1 MATERIALS
The TME 419 cultivar (Manihot esculanta) used for this study was obtained
from National Root Crops Research Institution (NRCRI) Umudike with the
maturity of about 6 months, Fifty kilogram (50kg) of cassava was used for
this research work and were processed after six hours of harvesting. And
also the soybean in this study was purchased from ekeonunwa market
owerri and processed into malted soyflour (fig 2)

3.2

PREPARATION OF MALTED SOYBEAN EXTRACT

FORTIFIED (FORTIFIED GARRI) GARRI

The fortified gari was produced at National Root Crop Research Institute
Umudike. The cassava tubers were manually cleaned by removing the back
by peeling. The peeled cassava tubers were washed properly and grated.
After which the cassava mash was divided into two, one part was used as
the control while the second part of the grated cassava mash with malted
soy-extract grated cassava mash at a ratio of 1300g of cassava mash was
mixed with 114.15g malted soy floor for 18% (w/w) soy fortification. The
control sample (white garri) had its cassava mash after measuring they were
then differently bagged and were allowed to ferment/dewatered, one was
mixed with soy-flour in the of 1kg of soy flour was added to 11.4kg of
30

cassava mash. Which was the ratio specified the white gari had its cassava
mash as 29kg after measuring they were then differently bagged and were
allowed to dewatered for 48 hours after which the sample was garrified
(toasted) and cooled in a sterile environment The gari was sieved through a
mesh size double layer muslin cloth to obtain a fine gari texture. The gari
was packaged in an air tight Ziploc bag.

3.3

SAMPLE PREPARATION FOR GARI (UNFORTIFIED

CONTROL)

The cassava tubers were peeled to remove the back, the peeled cassava were
washed and grated, after which the washed and grated cassava mash were
allowed to ferment alongside dewatering for 48hours, Dewatered/fermented
cassava mash were sifted and toasted (garrified) and cooled, the garri was
packaged in an air tight Ziploc bags.

3.4

MOISTURE VARIATION

250g of the fortified gari was separated into three different moisture levels
of 8%, 10% and 12% moisture.

31

3.5 MICROBIOLOGICAL DETERMINATION OF THE STORED

SAMPLES
Microbiological: -Twenty five grams proportion of each sample was
asceptically taken (after thorough mixing) and weighed into a beaker
containing 225ml of 0.1% sterile peptone water (w/v) and allowed to soak
for 2-3 minutes with occasional stirring with a sterile glass rod. Ten-fold
serial dilution was subsequently prepared by transferring 1ml aliquot of the
supernatant into 9ml sterile peptone water as diluents. Further serial dilution
was carried out and thereafter. The number of viable microorganism that
developed were counted and calculated and expressed as colony forming
units per gram (cfu/g).
Isolation and characterization. The fungi isolates were identified based on
examination of the conider heads, phialides, conidiophores and presence and
absence of foot cells or rhizoids

32

CASSAVA TUBER

WASHING

PEELING

WASHING

GRATINGING

FERMENTATION/DEWATERING

SIFTING

TOASTING (GARIFICATION)

COOLING

PACAKING

Fig 1: The flow diagram for the preparation of garri

33

SOYBEANS

STEEPING (10HRS)
SPROUTING (48HRS)
DRYING (600 C)
DEHULLING/WINOWING
MILLING

MALTED SOY FLOUR

Fig. 2: Flow diagram for the preparation of malted soy flour

34

3.4.1 Microbiological Media Preparation

Nine milliliters (9ml) of sterile distilled water was pipetted into a clean dry
test tube plugged with cotton wool and wrapped with aluminum foil. The
test tube were placed inside an autoclave and sterilized by autoclaving at
1210C for 15minutes. Nutrient agar (NA) Twenty eight grams (28g) of
powdered commercially prepared nutrient agar was weighted on analytical
mettle balance into clean dry one liter (1L) conical flask and 1000ml of
distilled water placed inside a water bath set at about 90c and allowed to
dissolve. it was then distributed into Mccartney bottles and sterilized in an
autoclave set 1210C for 15minites, macConkey Agar (MCCA) fifty-five
grams (55g) of MacConkey Agar was weighed into one liter capacity of
conical flask and brought to boil to dissolve the agar. It was then distributed
in MacConkey bottles and autoclave as for nutrient agar, Potato Dextrose
Agar (PDA) thirty-nine grams (39g) of PDA was weighed into a one liter.
Capacity of conical flask bring to boil and distribute them into MacConkey
bottles and autoclave as for nutrient agar, sample preparation The sample
(10g) was suspended in a 90ml of sterile distilled water and was
homogenized. The suspension was filtered through sterile wool. The
samples were serially diluted under aseptic condition.

35

CHAPTER FOUR
4.0 RESULTS AND DISCUSSION

4.1

MICROBIAL COUNT ON CASSAVA GARRI

Fig. 4.1: Effect of moisture variation on the total bacteria count of garri
samples

36

4.1.1. Effect of moisture variation on the total bacteria count of garri

during storage.
It was observed from this study that the total bacterial count of the garri
samples having different moisture levels (8%, 10% and 12%) stored at
ambient temperature condition showed a progressive increase in the total
plate count for aerobic mesosphillic bacteria all through the ten weeks
storage time (Fig. 4.1). From the graphical presentation, the fortified garri
samples had more total bacterial count compared to the control garri
samples which had lesser bacterial growth through the period of srorage. It
can also be seen that there is a significant increase in the aerobic mesophillic
bacterial growth for week 8 and week 10, this could be due the increase in
metabolic activities of microbes during the period of storage. The growth
rate shown in week 4 and week 6 are the insignificant. The total viable
bacterial count increased from week 0 3.8x103, 4.2x 103, 28.9x103, 31.0 x
103, 46.7x103 to 53.0x103 cfu/g for control 1 at the same moisture storage
content.

37

Fig.4.2: Effect of moisture variation on the total fungi count of the garri
samples

38

4.1.2. Effect of moisture variation on the total fungal count of garri

during storage
It can be seen from Fig 4.2, there were no fungal growth at week 0 for both
the control and fortified sample, at week 2, there were still no growth in the
control sample and insignificant fungal growth with the fortified samples
increasing as the moisture increases. Week 4 and week 6 showed the same
level of growth while a higher total fungal growth was shown in week 8 and
week 10. The total viable fungi count increased from about 18.2 x 10 3 cfu/g
from week four (4) to 32.1 x 103 cfu/g at week ten(10).

39

Fig. 4 3: Effect of moisture variation on the yeast/mould growth on the

garri samples

40

4.1.3. Effect of moisture variation on the yeast/mould growth of garri

during storage.
From fig 4.3, appearance of yeast/mould started at week 4 and week 6
almost having the same reading. There was an increased in mould /yeast
count from 5.9 x 103 cfu/g from week four (4) to 16.9x103 cfu/g at week 10
all for occurrence in all control samples and all soy fortified samples
respectively. The steady and gradual increase in yeast and mold growth is
regarded as the total viable yeast and mould counts present in all the
samples stored at different initial moisture content suggests a favorable
micro environmental condition and nutrients availability. The results of the
table 4.3 shows that the soy fortified garri stored at an initial moisture
content of 10% exhibited has higher yeast and mould stability with higher
microbial count of 22.5 x 103 cfu/g and bacterial at 56.13 x103 cfu/g and the
lowest yeast/mould count at 6.9 x103 cfu/g the result compared of the soyfortified garri and the ordinary white garri which serves as control at the end
of 10 weeks.

41

Table 4.1:

Fungi (Mould/Yeast) species isolated from garri samples

after ten weeks of storage.

Fungi

Samples
Control garri

Fortified garri

Aspergillus spp

Penicillium spp

Rhizopus spp

Geotrichum spp

Cladosporium spp

Rhodotorula spp

42

4.1.4. Fungi (Mould/Yeast) species isolated from garri samples after ten
weeks of storage.
Table 4.1, shows that Aspergillus spp, Penicillium spp, Cladosporium spp
and Rhodotorula spp were isolated from the control sample after 10 weeks
of storage at varying moisture levels while Rhizopus spp, Cladosporium
spp and Rhodotorula spp were detected and isolated from fortified garri at
different moisture levels. This result shows that fortified garri samples has
lower fungal incidence compared to the control. This could be attributed to
the high protein content in the fortified samples..

43

Table 4.2: Bacterial species isolated from garri samples after ten
weeks of storage.
Bacteria

Sample
Control garri

Fortified garri

Bacillus spp

Staphylococcus spp

Pseudomonas spp

Micrococcus spp

Acinetobacter spp

Achromobacter

44

4.1.5. Bactereria species isolated from garri samples after ten weeks of
storage.
From table 4.2, the two samples shared the same bacterial species which
includes; Staphylococcus spp, Pseudomonas spp, Micrococcus spp,
Acinetobacter spp and Achromobacter spp. after 10 weeks storage except
for the incidence of Bacillus spp isolated from fortified garri due to the high
protein content of the malted soy fortified sampleswhich was absent in the
control sample.

45

Table 4.3: Mean value of the total bacteria count from garri produced
TIME (WEEK)

Cfu/mg x 104

5.12d1.23

5.53d1.20

30.35c1.43

32.57c1.06

47.99b1.12

10

56.13a2.44

LSD

2.95

ad=Means with different superscripts along the columns differ significantly at P< 0.05.
LSD - Least significant difference

4.1.6. Mean value of the total bacteria count from garri produced
According to table 4.3 above, there exists significant difference in the total
bacterial count

46

Table 4.4: Mean value of the total Mould and Yeast count from garri
TIME (WEEKS)

Cfu/mg x 104

00

00

6.9c0.79

7.42c1.08

9.65bc1.47

10

22.05a4.38

LSD

4.01

ac Means with different superscripts along the columns differ significantly at P< 0.05.
LSD - Least significant difference

Table 4.5: Mean value of the total Fungi count from garri produced
TIME (WEEKS)

Cfu/mg x 104

00

0.68d0.79
47

20.12c1.42

21.28c1.62

28.45b0.80

10

33.47a0.87

LSD

2.16

ad Means with different superscripts along the columns differ significantly at P< 0.05.
LSD - Least significant difference

48

CHAPTER FIVE
5.0 CONCLUSION AND RECOMMENDATION
The storage quality of this garri therefore depends on the rate of
reproduction and growth of these organisms which in turn depends on some
biological and non-biological variable. The most important of this variable
is the storage moisture content which invariably guarantees the survival of
any of these organisms in any stored product.
It can therefore be concluded that moisture content is an important factor to
be considered in establishing the shelf life stability of cassava flour.
Moisture content had significant effect on the storage time as it was
carefully observed in the cassava garri. The moisture content of cassava
garri also had significant effect on the microbial content of the garri during
storage. Lower moisture content in the garri especially the fortified garri
corresponds to lower microbial growth and hence longer shelf-life stability
and better quality attributes. Low moisture content also resulted in higher
dry matter content, which is a good attribute for the food industries and
cassava garri processing. Cassava garri sample B( soy fortified garri) stored
in Ziploc bag at an initial moisture content of 0 10% provided the lowest
moisture content of 0 10% and the maximum stability of microbial quality
of cassava garri during the ten weeks storage period. The garri sample B
49

showed the highest bacteria content of 56.13 15% and the

highest fungi

counts of 33.47 15% at the end of ten weeks storage period. Cassava garri
control sample stored at an initial moisture content of 5.12 at 10% had the
lowest percentage loss in However, based on the overall quality and
microbial safety, a moisture content of 0.68 12% would be recommended
for long term storage of cassava garri in terms of quality attributes, shelf life
and microbial stability all of which are essential to food industries.

5. 1 RECOMENDATION
From all the observation the microbial test carried out on the two different
garri samples (fortified and the control) its advices that any processed garri
is best consumed within the range of one month of production and must be
kept under room temperature storage is best effective in an air tight
packaging material like zip-lock bag to avoid intrusion of environmental
organisms.

50

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Removal. Public lecture delivered at the Rivers State University of
Science and Technology. Port Harcourt.
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paste viscosity of cassava flour on fried cassava chips quality J. Food
Sci. and Agric. 42:67-7.
Akoroda, M.O. and Arene, O.B. (1989). Tropical Root Crops. In
proceedings of the Fourth Triennal of the International Society for
Tropical Root Crop, Africa Branch held in Kinshasa Zarie 5-6,
December pp198-201.
Balagopalan, C., Padnaja G., Nanda, S.K and Moorthy, S.N. (1988).
Cassava Starch In: cassava in Food, Feed Industry. CRC Press, Inc.
Boca Ratou, Florida. Pp113-126.
Dufor, D.L. (1989). Effectiveness of Cassava Detoxification Technique used
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51

Edem, D. O., Ayatse, J.O.I. and Itam . E.H (2001). Effect of soy protein
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Harbor, C.I. and Ogundu, E.C. (2009). Effect of processing on cyanide

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IITA, (1990). Cassava in the Tropical Africa. A reference manual (edited)

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1996

Microorganisms

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Foods

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Kwatia, J.T. (1986). Report on the Existing Cassava Storage and

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view of making recommendations for the establishment of rural
cassava

processing

and

utilization

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UNICEF/IITA

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Macrae, R. Robinson, R.K. and Sadler, M.G. (1993). Cassava. In

Encyclopedia of Food Science, Food Technology and Nutrition.
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Olaniyan J (1991). Microbial and food analysis of locally produce garri

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55

APPENDIX 1

Anova: Single
Factor

total bacteria count

SUMMARY
Groups

Count

Sum

Average

Variance

SD

week 0

30.7

5.116666667

1.513666667 1.230312

week 2

33.2

5.533333333

1.430666667 1.196105

week 4

182.1

30.35

1.307 1.143241

week 6

195.4

32.56666667

1.130666667 1.063328

week 8

287.9

47.98333333

1.257666667 1.121457

week 10

336.8

56.13333333

5.942666667 2.437759

ANOVA
Source of

Variation

SS

df

Between Groups

13379.79139

2675.958278 1276.055024

Within Groups

62.91166667

30

2.097055556

Total

13442.70306

35

56

MS

P-value
5.54E-

cri

34 2.533

Anova: Single
Factor

yeast and mould CFU

SUMMARY
Groups

Count

Sum

Average

Variance

SD

Week 0

Week 2

Week 4

41.4

6.9

0.62

0.787401

Week 6

44.5

7.416666667

1.181666667

1.087045

Week 8

57.9

9.65

2.147

1.465264

Week 10

132.3

22.05

19.195

4.38121

ANOVA
Source of
Variation
Between
Groups

SS
1974.118056

df

MS

5 394.8236111 102.3580965
57

P-value

F cri

6.41E-18 2.53355

Within Groups

115.7183333 30 3.857277778

Total

2089.836389 35

Total
Anova:

Fungi

Single Factor

Count
58

SUMMARY
Groups

Count

Sum

Average

Variance

Week 0

Week 2

4.1

0.683333333

0.681666667

0.7

Week 4

120.7

20.11666667

2.009666667

1.4

Week 6

127.7

21.28333333

2.633666667

1.6

Week 8

170.7

28.45

0.643

0.8

Week 10

200.8

33.46666667

0.758666667

0.8

ANOVA
Source of
Variation
Between

SS

df

Groups

5909.286667

Within Groups

33.63333333 30

Total

MS

P-value

1181.857333 1054.183944

9.5631E-33 2.5335

1.121111111

5942.92 35

tbc

Treatments
SAMPLES WK 0

WK 2

WK4

WK6

WK 8

WK 10

CON 1

3.8

4.2

28.9

31

46.7

53

CON 2

4.1

4.7

29.4

31.8

47.3

54.2

59

Fc

CON 3

4.3

4.8

29.9

32.4

47.1

54.8

SOY 1

5.7

5.8

30.8

32.9

48.5

57.7

SOY 2

5.9

6.3

31.2

33.4

48.7

58.2

SOY 3
6.9
7.4
Key: TBC = Total Bacterial Count

31.9

33.9

49.6

58.9

Yeast & Mould

SAMPLES WK 0
CON 1
0
CON 2
0
CON 3
0
SOY 1
0
SOY 2
0
SOY 3
0

Treatments
WK 2 WK4 WK6 WK 8 WK 10
0
5.9
6.3
7.8
16.9
0
6.3
6.4
8.3
19.8
0
6.5
6.7
9.4
21
0
7.2
7.9
9.9
19.8
0
7.6
8.4
10.9
26.7
0
7.9
8.8
11.6
28.1

Total fungi count

SAMPLES WK 0
CON 1
0
CON 2
0
CON 3
0
SOY 1
0
SOY 2
0
SOY 3
0

Treatments
WK 2 WK4 WK6 WK 8 WK 10
0
18.2
19.4
27.2
32.1
0
18.9
19.8
27.9
33
0
19.6
20.4
28.4
33.6
0.8
21
22.1
28.7
33.4
1.4
21.4
22.8
29.3
34.1
1.9
21.6
23.2
29.2
34.6

60