MICROBIAL AND NUTRITIONAL QUALITY OF PLANTAIN.

(Musa parasidisiaca)

BY
CHINWO, LUCKY CHILE
U2016/5555218

A PROJECT SUBMITTED TO THE DEPARTMENT OF

MICROBIOLOGY, FACULTY OF SCIENCE
UNIVERSITY OF PORT HARCOURT

IN PARTIAL FULFILMENT OF THE REQUIREMENT FOR THE

AWARD OF BACHELOR OF SCIENCE (B.Sc.) DEGREE IN
MICROBIOLOGY

FEBRUARY 2023

1
 CERTIFICATION

This is to certify that this project was carried out by CHINWO, LUCKY CHILE, Student of the

Department of Microbiology in the Faculty Of Science of the University Of Port Harcourt,

Choba, Port Harcourt, Rivers State with Matric No: U2016/5555218.

Dr. (Mrs.) N.J.P Omorodion ………............... ……………….

Supervisor Signature Date

Prof. H.O Stanley ………………. .. ……...............

Head, Department of Microbiology Signature Date

Prof. E.B. Essien ………….............. ………………

Dean, Faculty of Science Signature Date

……………………………. ……………………. ………………..

External Examiner Signature Date

2
 DEDICATION

This project is dedicated to God Almighty, the giver of wisdom, knowledge and understanding

and sustainer of life, for His unconditional love and mercy granted to me throughout the period

of this project. The research work is conspicuously dedicated to my family and to the world of

academia.

3
 ACKNOWLEDGEMENTS

I give thanks to Almighty God, who gave me the gift of life, and made this project a success. My

special thanks also go to my amazing supervisor Dr Mrs N.J.P Omorodion for accepting me to be

under her supervision and for her timely words of encouragement, instructions, and motherly

love.

This acknowledgement would be incomplete if I fail to express my gratitude to my family most

especially my mom, Mrs. CHINWO CHARITY, for her prayers, and financial support to ensure

my success

My special thanks also go to Prof. Mrs O.K. Agwa for her timely words of encouragement.

Special thanks to Madam Felicia for accepting me to work at the Laboratory.

Furthermore, I also want to use this opportunity to thank the laboratory staffs for their patience

and show of love throughout my stay in the laboratory. May God bless you all abundantly.

4
 ABSTRACT

Plantain (Musa parasidisiaca) is a major food crop in the humid and sub-humid parts of Africa
and major source of energy for millions of people in these regions. It is a perennial crop that
grows well in a wide range of environments and belongs to the family of Musaceae with the
genus Musa and have been crops of extraordinary significance to human societies. This study
was conducted to evaluate the microbial and nutritional quality of plantain sold in Port Harcourt.
A total of 30 samples of plantain comprising boiled, roasted and raw plantain. Microbial analysis
was done using standard microbiological methods. nResult obtained shows the bacterial load
level of different plantain samples that ranged from 0-3.62 x10 4 cfu/g for Total heterotrphic
count, 0-3.2 x104 fu/g for total staphylococcus count and 0-4.3x103 cfu/g for total coliform
counts. Fungi ranged from 0-2.5 x106 cfu/g respectively. The proximate test shows the
nutritional and mineral composition of plantain. 40% Staphylococcus sp 15% Aspergillus sp,
amd 14% fusarium sp etc were present in the all the samples. Other isolates include Bacillus sp,
mucor sp and Yeast sp etc. The proximate tests yielded a value of 3.50-5.14 for protein, 30.33-
47.42 for carbohydrate, 36.66- 58.24 for moisture in ripe and unripe. This shows evidence of
high nutritional value of plantain and lesser likelihood pf unripe plantain contamination. The
pattern of distribution showed that most of the isolates found in ripe and unripe plantain were
obtained from hawkers and roasted plantain (bole). These results imply that the government
should call for closer monitoring and control of the environment where roadside foods are
processed and sold.

5
 TABLE OF CONTENTS
Title page……………………………………………………………………………………. i
Certification............................................................................................................................ ii
Dedication…………………………………………………………………………………... iii
Acknowledgements……………………………………………………………………….... iv
Abstract ………..…………………………………………………………............................ v
Table of contects…….…………………………………………………………………….....vi
List of Table…………………………………………………………………………….…...vii
List of Figures………………………………………………….………………………..…..viii
CHAPTER ONE
1.0 1.1 …………………………………………………………………….………… 1
1.2 ……………………………...……………………………………………….. 1
1.3 …………………………………………………………......…………………1-2
1.4 ……………………………………………………………………………… 1
1.5 ………………………..………………………………………………………..4

CHAPTER TWO
2.0 2.1 ………………………………………………………………….………….... 5
2.2 ……………………………………………………………………………… 5-6
2.3 ………………………………………………………………………………. 5
2.4 …………………………………………………………………..…………… 5
2.5 ………………………………………………………………………..…...…. 5
2.6 ……………………………… ………………………………………………..7
CHAPTER THREE
3.0 3.1 ………………………………………..….…………………………………… .8-10
3.2 ……………………………………..………………………………………..… 10-12
3.3 …………………………………………………………………………….….... 12-21
3.4 …………………………………………………………………………..……... 21-23
3.5 …………………………………………………………………………………. 23-25
3.6 ………………………………………..……………………………………….... 25-26
CHAPTER FOUR
4.0
4.1 Discussion and Conclusion…………………………………………………
4.2 Recommendation
4.3 References
4.4 Appendix

6
 LIST OF TABLES

TABLE 4.1 Total Heterotrophic Bacteria Counts of Ripe and unripe plantain pulp samples

TABLE 4.2 Total Heterotrophic Bacteria Counts of Ripe and Unripe Roasted plantain

samples

TABLE 4.2 Total Heterotrophic Bacteria Counts of Ripe and Unripe Boiled plantain a

samples

TABLE 4.3 Total Staphylococcal Counts of Ripe and unripe plantain pulp samples

TABLE 4.4 Total Staphylococcus Counts of Ripe and Unripe Roasted plantain a samples

TABLE 4.4 Total Staphylococcus Counts of Ripe and Unripe Boiled plantain a samples

TABLE 4.5 Total Coliform Counts of Ripe and unripe plantain pulp samples

TABLE 4.5 Total Coliform Counts of Ripe and Unripe Roasted plantain a samples

TABLE 4.5 Total Coliform Counts of Ripe and Unripe Boiled plantain a samples

TABLE 4.5 Total Fungal Counts of Ripe and unripe plantain pulp samples

TABLE 4.5 Total Fungal Counts of Ripe and Unripe Roasted plantain a samples

TABLE 4.6 Total Fungal Counts of Ripe and Unripe Boiled plantain a samples

TABLE 4.9 Biochemical Characteristics of Bacteria Isolated from Ripe and unripe plantain

pulp samples

TABLE 4.10 Biochemical Characteristics of Bacteria Isolated from Ripe and Unripe Roasted

plantain a samples

TABLE 4.10 Biochemical Characteristics of Bacteria Isolated from Ripe and Unripe Boiled

plantain a samples

7
TABLE 4.11 Cultural/Cell Morphology and identification of Fungal Isolates obtained from

Plantain samples

TABLE 4.12 Proximate composition of Ripe and Unripe plantain pupl

8
 CHAPTER ONE

1.0. INTRODUCTION AND LITEARATURE REVIEW

1.1. INTRODUCTION

1.1.1 BACKGROUND OF THE STUDY

Plantain (Musa parasidisiaca) is a major food crop in the humid and sub-humid parts of Africa

and major source of energy for millions of people in these regions. It is a perennial crop that

grow well in a wide range of environments (Nelson et al., 2006), and belong to the family of

Musaceae with the genus Musa and have been crops of extraordinary significance to human

societies.

Presently, it ranks as the fourth most important food crop in the world after rice, wheat and

maize; and are used as food, beverages, fermentable sugars, medicines, flavourings and

cooked foods (Nelson et al., 2006; Phillip et al.,2009). Plantain plant consists of long,

overlapping leafstalks and bears a stem which is 1.22 to 6.10 m high and produces bunches with

fewer but bigger fingers, and it is used locally in various forms by humans. The total

production of plantains in 1988 was 24.0 million metric tonnes (Food and Agriculture

Organization of the United Nations (FAO, 1988)), and in Nigeria, plantain production is

estimated at about 2.4 million metric tonnes mostly obtained from the Southern state (Folayan

and Bifarin, 2011). This accounts for its wide use in diverse ways alongside other foods as

staple in Nigeria. Plantain tends to be firmer and lower in sugar content. Plantain is normally

cooked or process, and are used either when green, under-ripe or over-ripe. Half-ripe plantai

is usually processed into plantain flour by slicing the plantain and sun drying for some days

and cooked into sticky paste delicacy “Amala ogede” (Yoruba), and “Ebue” (Ogonis)

served with vegetable soup. Half-ripe plantain is also boiled, fried, processed into chips.

9
Boiled and pounded plantain pastry are eaten with soups, sauce or vegetables. Ripe plantain

flour has been used in making bread, biscuits and instant flour. The nutritional qualities and

sensory attributes of wheat bread substituted with 15% plantain flour were com-parable to

that of whole wheat bread; hence, its adoption was recommended in bread making processes.

The Soyamusa, a baby food from plantain flour (60%) was made and used in Nigeria. In

Nigeria, as one of the major staple foods, plantain is processed into various products such as

‘elubo’ (dried half-ripe plantain flour), ‘dodo’ (fried sliced ripe plantain pulp), roasted

plantain (bolle), chips (fried half-ripe pulp), and in addition to yam, it can be pounded to a sticky

paste eaten with soup. It can also be processed to food/foodstuffs such as breakfast cereals,

baby complementary foods. The International Institute for Tropical Agriculture (IITA, (2005)

reported that post-harvest loss of plantain is one of the major threats to the availability of the

fruit that is a staple to many Nigerians. Some research was made and reported that fermentation

may impart new colour, flavour, taste, and texture to food products, as well as enhancing the

nutritive value and extending the shelf-life of the fermented products.

The current trend in nutrition in meeting consumers’ daily dietary needs is promotion of

dietary diversification through locally available foods. However, little is known about the

nutrient composition and nutrient retention of processed plantain products. It is therefore the

objective of this study to provide information on the nutrient composition and potential

contribution of unripe, ripe, boiled and roasted plantain to dietary diversity of Nigerian

consumers.

Although, it has been found that microorganisms are seen in the raw samples of plantain with

high counts and presence of Bacillus and staphylococcus species amongst others which can lead

10
to serious health risks if not properly prepared before use. Production methods and techniques

vary from one locality to another resulting in a product of variable quality indices and shelf life.

Also, some fungi discovered in plantain samples cause rot and destruction of produce which

have economic effects. Lost produce cannot be consumed or sold in the market anymore. Black

sigatoka (Mycosphaerella fijiensis) is a fungal disease that affects production of plantain crops in

tropical regions.

1.1.2 STATEMENT OF PROBLEMS

Plantain is a widely consumed staple food in the African sphere. Due to its popularity, it has

been processed and eaten in a variety of forms. Some of these include boiled, roasted, fried, dried

and even milled plantain flour. During the processing, the plantain products are easily

contaminated which can lead to food poisoning when consumed. Hence, it is imperative that the

microbiological quality of all the varieties of processed plantain be determined to come to a

conclusion of a best method of preparation and also areas for caution in consumption. In this

study we took into consideration the microbiological quality of boiled, roasted and raw ripe and

unripe plantain.

1.1.3 AIM OF RESEARCH

The aim of this research is to determine the microbial and nutritional quality of plantain (musa

parasidisiaca).

1.1.4 OBJECTIVES OF STUDY

The objectives of this study were:

To determine the following microbial analysis:

11
 i. To attain total bacteria and fungi count

ii. To attain total coliform and total staphylococcus count.

iii. The pH of the samples will be checked on each stock sample.

iv. To determine the proximate composition of ripe and unripe plantain

1.1.5 SIGNIFICANCE OF THE STUDY

This study is done to find out the microbial and nutritional quality of ripe, unripe, boiled and

roasted plantain (musa paradisiaca). Also, this study gives an insight into the conditions as well

as factors that result in ripe, unripe, boiled and roasted plantain, not to mention the nature of

microbial analysis or group of microorganisms that are associated with such a food.

The significance of this study is seen in the light of having a good understanding and showing

the microbial and nutritional composition of ripe, unripe, boiled and roasted plantain

This work will serve as a dependable source of information to food industries.

This research project will also be useful in the control and prevention of microorganisms

associated with food products like plantain (musa parasidisiaca).

1.2. LITERATURE REVIEW

Plantain belongs to the genus Musa of the family musaceae. Nearly all edible plantain cultivar

are derived from two wild species, M. acuminate and M. balbisiana (Robinson, 1996). These

wild species are classified on the basis of the proportion of the genetic constitution contributed

by each parental source (Robinson, 1996). Plantain is a staple crop and an important dietary

source of carbohydrate in Nigeria and in the humid tropical zones of Africa, Asia and South

America (Robinson, 1996). Plantain is rich in vitamins A, C and B group as well as minerals

such as calcium and iron (Marriott and Lancaster, 1983; Robinson, 1996). Plantain provides

12
between 9% and 35% of the total calories in the diets of more than 14 million people in Sub-

Saharan Africa (Robinson, 1996). The contributions of this staple starch crop to the food chains

of this region cannot be overemphasized (Robinson, 1996). Plantains are typical climacteric

fruits in that they exhibit a well-defined pre-climacteric phase after harvesting during which the

fruit remains unripe, the basal respiration rate is low and ethylene production is almost

undetectable. The respiratory climacteric commences spontaneously and there is a rapid and

well-defined rise in respiratory rate which is closely synchronized with evolution of ethylene,

with chlorophyll breakdown in the peel and with starch to sugar conversion and tissue softening

in the pulp (Marriot and Lancaster, 1996). The fruit usually harvested at its mature but unripe

stage, ripens within two to seven days, thus making plantain a highly perishable crop,

particularly in the overripe stage (Robinson, 1996). An unripen plantain has high starch and low

sugar levels plus copious amounts of bitter tasting latex. Starch is converted to sugar as the fruit

ripens. (Daniells et al., 2001) FAO (2004) data sources put the world production of plantains at

about 60 million tons (FAO, 2004). In West Africa, plantain production increased at an average

annual rate of between 2.3% to 2.6% (FAO, 2004). The level of production of plantains in Africa

is comparable with other fruits like grapes (57 million tons); citrus (50 million tons) but much

greater than most other important fruits like apples (21 million tons) and mangoes (13 million

tons) (FAO, 2004). The higher production figures for plantains has been attributed to the cheaper

methods of growing that require few labor inputs, little soil preparation and little weeding are

needed once the plant has established vegetative cover. (FAO, 2004).

13
1.2.1 MATURITY INDICES OF MUSA sp.

Plantain requires about three months from the beginning of flowering until harvest. Multiple

fruits are produced on a large bunch, weighing between 50-200kg. Within the bunch are clusters

of double rows of fruit called “hands” and individual fruit called “fingers”. Maturity standards

for plantains are less precise. Several different external and internal fruit characteristics can be

used to determine plantain maturity. These include fruit diameter, age of the bunch, angularity of

the fruit, length of the fruit, and peel color (Johnson et al.,1998). The stage of maturity for

harvest depends on the intended market destination (Johnson et al.,1998). Locally marketed

plantains can be harvested at a more advanced maturity stage compared to export market fruit.

Export market destined fruit should be harvested the day before or the same day of shipment.

Plantain maturity is related to the diameter of the fingers. This is determined by measuring the

diameter of the fruit at its midpoint with a pair of calipers. Another method for estimating

plantain maturity is to record the age of the bunch. The time from when the fruit bunch first

becomes visible (Shooting) is recorded. Bunches can be tagged with different colored ribbons at

the time of shooting, and subsequently harvested after the appropriate time for the particular

cultivar, based on the season of the year and experience (Johnson et al.,1998). The colour of the

ribbons is changed weekly to coincide with the time of shooting and subsequently the age of the

bunch (Johnson et al.,1998). A third method used to determine harvest maturity is to observe the

shape (fullness) and angularity of the fruit. Immature fruit is angular in cross-sectional shape and

has distinct ridges. As the fruit matures, it becomes less angular and more rounded or full. The

degree of roundness differs between cultivars and location of the hand on the bunch. Typically,

the fullness of the fruit on the middle hand is measured. The appropriate shape to harvest the

14
fruit depends on the market destination. Fruit intended for the domestic market should be

harvested when the fruit shape is nearly round (Johnson et al., 1998).

A fourth way of estimating plantain bunch maturity is to measure the length of the edible pulp

portion of the fruit from the fingers in the middle hand. The length should be a minimum of

15cm for the domestic market and 18cm for the export market (Johnson et al.,1998). Finally,

peel colour is another frequently used method of assessing fruit maturity.The peel remains green

throughout growth and development of the fruit until it reaches physiological maturity. It then

changes to a yellow colour during ripening. However, plantain fruit should be harvested when

the peel is green in colour to withstand the rigors of handling and distribution. Internal fruit

composition changes dramatically during plantain fruit ripening. At physiological maturity, the

fruit is fully developed in size, green in peel colour, and at its highest level of starch. The starch

will progressively be converted to sugar as ripening progresses. The stage of harvest maturity of

plantains will depend on the target market. Plantains for local market are harvested at a more

advanced stage of maturity than those for exportation. However, if the fruit is too mature at

harvest, particularly following irrigation or rainfall, fruit splitting can occur during handling.

Also, mature fruit may ripen prematurely during transport or storage.

1.2.2 PROCESSING QUALITY

The bulk of the plantain are eaten either as raw, in the ripe state, or as a cooked vegetable, and

only a very small proportion are processed in order to obtain a storable product. Generally,

preserved products do not contribute significantly to the diet of the millions of people who eat

plantain, however in some countries or areas, the processed or preserved products are important

15
in periods when food is scarce. Processing is recognized as a way of preserving the fruit. Yet the

proportion of fruits processed and the suitability of the various Musa groups to processing is

relatively unknown. New Musa Therefore be screened for their processing quality or suitability

for processing pies (Adeniji et al.,2006). Ripe plantains are often sliced lengthwise, baked or

boiled, and served (perhaps with a garnish of brown sugar or chopped peanuts) as an

accompaniment for ham or other meats. Ripe plantain may be thinly sliced and cooked with

lemon juice and sugar to make jam or sauce, stirring frequently during 20 or 30 minutes until the

mixture jells. Whole, peeled plantain can be spiced by adding them to a mixture of vinegar,

sugar, cloves and cinnamon which has boiled long enough to become thick and then letting them

cook for 2 minutes, and they are mostly eaten with stew. Through experimental work with a view

to freezing peeled, blanched, sliced green plantain, it has been found that, with a pulp-to-peel

ratio of less than 1:3 the fruits turn gray on exposure to air after processing and this discoloration

is believed to be caused by the high iron content (4.28p/m) of the surface layer of the flesh. Its

reaction to the tannin normally present in green plantains. Plantain for freezing should have a

pulp content of at least 60% for maximum quality in the ultimate food product, but a range of 55

to 65% is considered commercially acceptable. In Ghana, plantains are consumed at 5 different

stages of ripeness (Chandler, 1995). Fully ripe plantains are often deep fried or cooked in various

dishes. A Ghanian pancake called “fatale” is made of nearly full ripe plantains and fermented

whole meal dough of maize, seasoned with onions, ginger, pepper and salt, and fried in palm oil.

“Kaklo” is the same mix but thicker and rolled into balls which are deep-fried. Because home

preparation is laborious, a commercial dehydrated mix has been developed. In Ghana, green

plantains are boiled and eaten in stew or mashed, together with boiled cassava, into a popular

plastic product called “fufu” which is eaten with soup. Because of the great surplus of plantains

16
in summer, technologists have developed methods for drying and storing of strips and cubes of

plantain for house use in making “fufu” out of season. The cubes can also be ground into

plantain flour. Use of infra red, microwave, and extrusion systems have resulted in high-quality

finished products. Processing has the added advantage of keeping the peels at factories where

they may be converted into useful by-products instead of being added to the bulk of household

garbage (Chandler, 1995). Plantain flour, or powder, is made domestically by sun drying slices

of unripe fruits and pulverizing (Anon, 1999). Commercially, it is produced by spray-drying, or

drum-drying, the mashed fruits (Anon, 1999). The flour can be mixed 50-50 with wheat flour for

making cupcakes. Two popular Puerto Rican foods are “pasteless” and “alcapurais” both are

pastry stuffed with meat, the first is wrapped in plantain leaves and boiled the latter is fried. The

pastry is made of plantain flour or a mixture of plantain with cassava or cocoyam. Commercial

production and marketing of fried green plantain and banana chips has been increasing in various

parts of the world over the past 25 years and these products are commonly found in retail

groceries alongside potato chips and other snack foods.

1.2.3 FLOUR AND POWDER

Flour can be made from green unripe plantain. Fruits are hand-peeled and sliced or chopped into

pieces about 5-10 mm thick. The slices will be dried in the sun by spreading out the slices on

mats, on bamboo framework, on cement floors, or on a roof or sheets of corrugated iron or

simply on a swept bare ground. Various designs of solar dryers can also be used, or they may be

dried in ovens, over fires, in a cabinet dryer or tunnel dryer (Thompson, 1995). The fruits are

either sun-dried which is the former, oven-dried, the latter or foam mat dried which will be

described now. Sun and oven-drying methods have been used for drying of plantain (Bowrey et

al.,1980; Johnson et al., 1998; Demirel & Turhan, 2003) with some success, the introduction of

17
foam-mat drying brought much more (Falade and Olugbuyi, 2009). In foam-mat drying plantain

puree was prepared by blending steam blanched plantain and distilled water for 2 mins in a

Waring blender to produce a 30 ± 0.4% total solids (TS) paste. A 20% (w ⁄ w) glyceryl

monostearate (GMS) suspension is prepared by dissolving a known weight of GMS in hot water

at 100oC. The 20% suspension is added to obtain a 0.02% GMS in the plantain paste. The

mixture of plantain paste (30% TS) and GMS suspension are then transferred into a Kenwood

Chef mixer and whipped at maximum speed for 4 mins until homogenous foam is obtained. The

whipped foam could be extruded using a manual Euroline icing syringe (Model 5 Nozzles

stainless steel 19 cm, Euroline, Essex, UK)with an outlet orifice of 4 mm diameter on a stainless

steel wire mesh and dried in a cross-flow Gallenkamp Oven at 60oC for 45–90 mins. The dried

plantain is scraped offand packaged in low density polyethylene (100 μm) to prevent moisture

absorption (Falade and Olugbuyi, 2009). After drying, the chopped pieces have a moisture

content of about 5-10%. The dried pieces were ground and usually sieved to produce the flour.

The flour is packaged in moisture proof bags. The dried slices are stored and only converted to

flour when needed since the flour tends to lose its flavour rapidly or may absorb moisture

(hygroscopic) and become mouldy. Powder could be prepared from fully ripe plantain. Plantain

(Musa spp) may be processed into many products at different stages of physiological maturity;

unripe, ripe, overripe or in a number of ways such as frying, grilling, boiling and drying.

Moisture removal from plantain seems to be an appropriate and economical means of preserving

Musa spp, resulting in shelf stable and convenience products. Currently, unripe plantain flour is

being processed into a thick paste product known as ‘amala’ in the western part of Nigeria,

which is medically recommended for diabetic patient (Adeniji et al., 2006). Improved cultivars

of plantain and may provide high quality whole flour from the entire fruit for livestock feed,

18
which may eventually provide protein in human diet from consumption of meat and other

products of livestock (Thompson, 1995.). Such flour may be employed in traditional dishes for

human consumption based on their nutritional profiles. Although, there is need to investigate the

application of whole Musa flour in baking and confectioneries from the point of view of their

pasting properties but that notwithstanding it has recorded success when used in addition to the

conventional wheat flour. The use of entire fingers of plantain could be a rapid approach in flour

production with improved levels of nutrients, especially minerals, which are concentrated in the

peel (Izonfuo and Omuaru, 1988).

1.2.5 PULP CRUSHING METHOD

There were two pulp crushing methods used; pounding in a traditional mortar and milling with

the disc attrition mill. With the exception of sample J, all the other samples were pounded. The

size reduction method used influences the consistency of the paste produced (Fellows, 2000).

Size reduction of the senescent plantain pulp is essential so as to obtain similar particle sizes with

flour and all other ingredient used for effective mixing. The size reduction also affects the texture

of the final product (Fellows, 2009). The cleanliness of the mortar used may also compromise

the microbial safety of the product. According to Sinayobye and Saalia (2011), the main

challenges with the use of the disc attrition mill are microbial and chemical contamination. The

microbial load of the samples, however, may subsequently be reduced by the cooking (baking or

steaming) step.

1.2.6. OTHER USES

Plantain leaves are widely used as plates and for lining cooking pits and for wrapping food for

cooking or storage. A section of leaf often serves as an eye-shade (Anon, 1999). In Latin

America, it is a common practice during rains to hold plantain leaf by the petiole, upside-down,

19
over one’s back as an “umbrella” or “raincoat” (Anon, 1999).. In West Africa, fiber from the

pseudostem is values for fishing lines. In the Philippines, it is woven into a thin, transparent

fabric called “agna” which is the principal material in some regions for women’s blouses and

men’s shirts. It is also used for making handkerchiefs. In Ceylon, it is fashioned into soles for

inexpensive shoes and used for floor coverings (Thompson, 1995). Plantain fibre is said to be

superior. The ash from the dried peel of plantains is rich in potash and used for making soap.

That of the burned peel of unripe fruits of certain varieties is used for dyeing.

1.2.7 MICROBIOLOGICAL CONTAMINATION

Microorganism associated with production and processing of plantain to obtain some valuable

end products includes Bacillus subtilis, Staphylococcus aureus, Lactobacillus spp.,

Saccharomyces cerevisiae, Aspergillus flavus, Aspergillus niger, Fusarium species and

Penicillium species (Fajinmi et al., 2011). Variable numbers of coliforms such as Escherichia

coli, Klebsiella pneumonia and Salmonella typhii, which may arise from unhygienic water,

materials and human contamination can also be present (Ohenhen et al., 2013; Kouadio et

al., 2014). Environmental factors that influence the growth of plantains include adequate

rainfall and temperature (28 -320C) which are ideal for the growth of the crop. Temperatures

below 180C and above will adversely affect growth. Stormy winds can also cause

considerable damage to plantain because of its week rooting systems (Li et al., 1999). Soil is

another major factor that affects plantains. Some nutrient components such as Nitrogen,

Phosphorus, Potassium, Magnesium and Calcium are the most important element for

plantains to avoid diseases conditions (Hosset al., 2000; Oloyede et al., 2013). Plantains and

Bananas can be affected by three major insect pests such as Plantain/Banana Root Weevil

– Cosmopolites sordidus; Pseudostem Borer –Lapaeumides licus(Lepidoptera; Castinidae) and

20
Fruit Scarring Beetle – Colapsishypochlora(Coleoptera; Chrysomelidae). Many microbial

agents are however involved in plantain infestation (Ruiz, et al., 2009).

21
 CHAPTER TWO

2.0 MATERIALS AND METHODS

2.1 Sample collection and preparation

Five samples of ripe and unripe plantain, ripe and unripe roasted plantain. Ripe and unripe boiled

plantain fruit were collected from different markets in Choba in Rivers state.

2.2 Sterilization of materials

A total of four media were used to isolate the microorganisms, the study for the isolation of both

bacteria and fungi. He media used are Nutrient agar, Sabouraud dextrose agar, MacConkey agar

and Mannitol salt agar. The various media, diluents and glass wares were properly sterilized in

the autoclave at a temperature of 1210C at 15psi for 15minutes

Preparation of serial dilutions

Aseptically 10 grams of the sample were homogenized in 90 ml of sterile diluent (0.1% Peptone

water). It was mixed well to give dilution (10-1) by using sterile pipette 1 ml was transferred

aseptically from dilution (10-1) to a test tube containing 1 ml of sterile diluent (10-2). One ml of

each dilution was transferred into sterile petri dish, and then 15 ml of sterile melted Plate Count

Agar medium were added to each plate. The inoculum was mixed with medium and allowed to

solidify. The plates were incubated at 37º C for 48 hours. A colony counter was used to count the

viable bacterial colonies after incubation and the results were reported as colony-forming units

(CFU) per gram.

22
2.3 Media preparation

A total of four different media were used for isolation of different microorganism from the

plantain samples. The media used include: Nutrient agar, Sabouraud dextrose agar, MacConkey

agar and Mannitol salt agar.

Nutrient agar

28g of agar powder was suspended in 1000 ml distilled water, heated to boiling to dissolve the

medium completely. The media was then sterilized in an autoclave at 121°C, at 15psi for 15

minutes, allowed to cool and was poured into petri dishes using aseptic techniques.

Sabouraud dextrose agar

65g of sabouraud dextrose agar powder into 1000ml of water was sterilized in an autoclave at

121°C, at 15psi for 15 minutes. The media was allowed to cool for 45 to 50°C and was poured

into petri dishes using aseptic techniques.

MacCkonkey agar

This is used for the isolation and differentiation of lactose fermenting and non-lactose fermenting

enteric bacteria. 101g of MacConkey powder was dissolved into 1000ml of distilled water. The

solution was sterilized by autoclaving at 121 0C for 15mins at 15psi. about 20ml of the solution

was dispensed on the sterile petri-dishes and allowed to solidify

Manittol salt agar

This medium is used for the isolation of Stapphylococcous sp. 111g of manittol salt agar was

dissolved into 1000ml of distilled water. The solution was sterilized by autoclaving at 121 0C for

23
15mins at 15psi. the media solution was dispensed on the sterile petri dishes for proper

solidification.

3.6.2 Peptone Water

0.1% peptone water was prepared by dissolving 1g of peptone powder into 100ml of distilled

water. The mixture was properly stirred with a glass rod before dispensing into test tubes for

autoclaving at 1210C for 15minutes at 15psi.

3.7 Preparation of Stock Culture

After sub-culturing of the isolates on nutrient agar, media agar slants were prepared in bijou

bottles by dispensing 10ml of nutrient agar solution into bijou bottles. The medium was sterilized

by autoclaving at 1210C for 15minmutes at 15psi. the bottles were slanted and allowed to

solidify, isolates from the pure cultures were inoculated on the surface of the agar slant and this

was incubated at 370C for 24hours.

2.4 Sample preparation

10g of each sample was transferred to a sterile conical flask with 90 ml of sterile peptone water

and homogenized and then serially diluted in sterile 0.1% peptone water.

2.6 Microbiological analysis

2.6.1 Total bacteria count (TVC)

Isolates were identified based on colonial morphology and cultural characteristics on growth

media which include: colony size, color, opacity, consistency, colony pigmentation elevation,

odour, swarming, identification materials, reagents and protocols were according to

Cheesbrough, 2005.

24
2.6 Total fungi count

This was done on the colonial morphology (colour, size and texture) and the cell morphology

(mycelium, hyphae) of the fungi using lactophenol blue. A piece of mycellium from the petri-

dishes was mounted on a clean grease-free slide using a sterile wire loop and was cove red with a

cover slip/ a drip of lactophenol cotton blue was added and allowed for few minutes before

examining under the microscope.

2.6.2 Total coliform count

The total coliform count was obtained using the Most Probable Number (MPN) method. The

presumptive, confirmatory and completed tests were carried out using MacCkonkey agar.

The presumptive test was done by adding 10ml of each sample onto 10ml of double strength

MacCkonkey agar in test tubes containing inverted Durham tubes. The setup was left for 24

hours at 37°C after which positive tube indicated by color change from purple to yellow and gas

production are subjected to a confirmatory test.

Using an inoculating loop, an aliquot from each of the first setup was transferred to a new setup

and left for 24 hours.

The completed test was carried out by inoculating freshly prepared EMB agar with cultures from

the positive tubes in the confirmatory test for 24 hours at the same temperature. The presence of

nucleated colonies with dark center is indicative of the presence of coliforms.

2.6.3 Total Staphylococcal count

1ml of each 10-3 diluted sample was spread on Mannitol salt agar plates, incubated at 25°C for

24-48 hours and observed for growth. The microbial organisms that show a characteristic colony

25
morphology of S. aureus on Mannitol salt agar (MSA) as a yellow-coloured sheen were

confirmed to be S. aureus. Further identification of S. aureus was done using the Gram staining

method, and standard biochemical assays such as oxidase and catalase tests.

2.7 Isolation and preparation of pure cultures

Distinct colonies were randomly collected using a sterile wire loop and sub cultured on fresh

sterile nutrient agar plates for bacteria and incubated for 24 hours for bacteria and sabouraud

dextrose agar and incubated for 5 days for fungi to obtain pure cultures.

2.8 BIOCHEMICAL TESTS

Catalase test

This test was used to detect the presence of catalase utilizing organisms which converts hydrogen

peroxide to water and oxygen. A drop of catalase reagent was placed on a grease free slide. One

loop full of the organism was placed on the catalase and observed for bubbling which shows a

positive result of catalase utilization.

Oxidase test

This was done to identify organisms that produce the enzyme cytochrome oxidase. Whatman

filter paper was moistened with oxidase reagent. A loop full of the organism was smeared on the

filter paper and observed for color change. A deep blue color shows a positive result.

Citrate test

This test was used to identify enterobacteria, based on the ability of the organism to use sodium

which converts hydrogen peroxide to water and oxygen as its carbon source. A medium of

Simmons-citrate agar was prepared accordingly. It was then boiled and dispensed into test tubes

26
and sterilized in the autoclave at 121°C at 15 Psi for 15 minutes. The organisms were then

stabbed into the medium and incubated for 48 hours. A color change from green to blue shows a

positive test.

Indole test

This test was done to differentiate gram negative rods. A 25g/ml peptone water solution was

prepared and sterilized. The solution was then dispensed into sterile test tubes. The test

organisms were inoculated into the solutions and incubated for 72 hours. After incubation, 0.5ml

of Kovac's reagent was added to each solution. A ring-like red coloration shows a positive test

for indole production.

Motility test

A half strength semi-solid medium having 14g/ml concentration of nutrient agar was prepared

and sterilized. The agar was then poured into sterile test tubes and allowed to cool and solidify.

The different colonies were collected using an inoculating needle and was stabbed into the

medium in different test tubes. The stab was incubated for 48 hours and observed for growth.

Dispersed growth from the line of stab shows that the bacterium is motile.

Sugar fermentation test

A bacteriological peptone solution was prepared 1%w/v of the sugars, in separate conical flasks.

Bromocresol purple reagent was then added as an indicator. The solution was dispensed into test

tubes with inverted Durham tubes.

The solution was autoclaved and allowed to cool before the solution was inoculated with the test

organisms and appropriately labelled. The set up was allowed for 72hours and results were taken.

27
The change of the solution from purple to yellow shows a positive result for sugar fermentation

and the formation of gas bubbles in the Durham tubes indicates gas production.

Triple Sugar Iron test

TSI agar was prepared accordingly, boiled and dispensed into test tubes.It was then sterilized and

allowed to cool in a slanted position to give a 2.5 cm butt and a 3.8 cm slant.. The different

colonies were collected using an inoculating needle and was stabbed into the center of the

medium to the bottom of the tube and then streaking on the surface of the agar slant in different

test tubes. The test tubes were incubated for 24 hours and observed for the production of gas and

colour change.

Methyl-Red test

An MR-VP medium was prepared accordingly. 9ml of the medium was dispensed into separate

test tubes. It was then sterilized and allowed to cool. A loopfull of the test organism was then

inoculated into the solutions. It was incubated for 48 hours. The test tubes were divided into two

sets. To one set, 3ml of 5% solution of alpha-naphtol was added. A red coloration shows a

positive result.

Voges -Proskuer test

An MR-VP medium was prepared accordingly. 9ml of the medium was dispensed

For Total Bacteria Count (TBC):

Nutrient agar

28
For Total Fungi Count (TFC)

Sabouraud dextrose agar (SDA)

For Total Coliform Count (TCC)

MacConkey agar

For Total staphylococcus

Manittol salt agar

into separate test tubes. It was then sterilized and allowed to cool. A loopfull of the test organism

was then inoculated into the solutions. It was then incubated for 48 hours.

The test tubes were divided into two sets. To one set, 1ml of 40% solution of KOH was added. A

pink coloration shows a positive result.

2.9 Gram staining

This test was carried out to determine the morphology of the cells and also determine if they are

gram positive or negative. Gram positive organisms appear purple, while gram negative

organisms appear red when viewed under the microscope.

A thin smear of the colony was made on a grease-free slide and heat-fixed. Crystal violet was

added to cover the smear for one minute and rinsed off with slow running tap water. Gram's

iodine was then added as a mordant to cover the smear for one minute and rinsed off. 95%

alcohol was added on the smear for 15 seconds and rinsed off. Safranin reagent was then added

to cover the smear for 30 seconds and washed off. The smear was allowed to dry and viewed

under the microscope.

29
2.10 Identification of fungi

The identification of the pure cultures of fungi was done by staining the fungi with lactophenol

cotton blue, viewing the specimen under the x40 objective lens of the microscope and using a

fungal atlas to identify the different pure cultures of fungi.

2.11 PROXIMATE COMPOSITION

Proximates are used in the analysis of biological materials as a decomposition of a human-

consumable good into its major constituents. They are a good approximation of the contents of

packaged comestible goods and serve as a cost-effective and easy verification of nutritional

panels. This means that testing can be used to verify lots, but cannot be used to validate a food

processor or food processing facility; instead, a nutritional assay must be conducted on the

product to qualify said producers. Nutritional panels in the United States are regulated by the

FDA and must undergo rigorous testing to ensure the exact and precise content of nutrients. This

should prevent food processors from making unfounded claims to the public.

In industry, the standard proximates are:

 Ash

 Moisture

 Proteins

 Fat

 Crude Fiber

 Carbohydrates (calculated)

30
The proximate compositions of ripe and unripe plantain pulp samples are shown in Table 4,

which was calculated on dry basis to aid comparison with literature data.

2.11.1 CHEMICAL METHODS

MATERIALS AND METHODS

2.11.1.1 MOISTURE CONTENT

The moisture content was determined according to the standard method of the Association of

Official Analytical Chemists (AOAC, 2003). Principle: The moisture content in a weighed

sample is removed by heating the sample in an oven (under atmospheric pressure) at 105 °C.

Then, the difference in weight before and after drying is calculated as a percentage from the

initial weight + 66.

Procedure: A sample of 2 g ±1 mg was weighed into a predried and tarred dish. Then, the

sample was placed into an oven (No.03-822, FN 400, Turkey) at 105 ± 1 °C until a constant

weight was obtained. After drying, the covered sample was transferred to desiccators and cooled

to room temperature before reweighing. Triplicate independent results were obtained for each

sample and the mean value was reported to two decimal points according to the following

formula:

Moisture content (%) = (Ws – Wd) × 100% Sample weight (g) Where: Ws = weight of sample

before drying. Wd = weight of sample after drying.

2.11.1.2 CRUDE PROTEIN CONTENT

The protein content was determined in all samples by micro-Kjeldahlmethod using a copper

sulphate-sodium sulphate catalyst according to the official method of the AOAC (2003).

31
Principle: The method consists of sample oxidation and conversion of its nitrogen to ammonia,

which reacts with the excess amount of sulphuric acid forming ammonium sulphate. After that,

the solution was made alkaline and the ammonia was distilled into a standard solution of boric

acid (2%) to form the ammonia-boric acid complex which is titrated against a standard solution

of HC1 (0.1N). The protein content is calculated by multiplying the total N % by 6.25 as a

conversion factor for protein.

Procedure: A sample of two grams (2 gm.) was accurately weighed and transferred together with,

4g NaSO4 of Kjeldahl catalysts (No. 0665, Scharlauchemie, Spain) and 25 m1 of concentrated

sulphuric acid ( No.0548111, HDWIC, India) into a Kjeldahl digestion flask. After that, the flask

was placed into a Kjeldahl digestion unit (No.4071477, type KI 26, Gerhardt, Germany) for

about 2 hours until a colorless digest was obtained and the flask was left to cool to room

temperature. The distillation of ammonia was carried out into 25m1 boric acid (2%) by using 20

ml sodium hydroxide solution (45%). Finally, the distillate was titrated with standard solution of

HC1 (0.1N) in the presence of 2-3 drops of bromocreasol green and methyl red as an indicator

until a brown reddish color was observed.

Crude Protein (%) = (ml Hcl sample – ml Hcl blank) x N x 14.00 x (F) ×100% Sample weight

(g) x 100 Where: N: normality of HCl. F: protein conversion factor = 6.25 20

2.11.1.3 FAT CONTENT

Fat content was determined according to the official method of the AOAC (2003).

Principle: The method determines the substances which-are soluble in petroleum ether (65-70

°C) and extractable under the specific conditions of Soxhlet extraction method. Then, the dried

32
ether extract (fat content) is weighed and reported as a percentage based on the initial weight of

the sample.

Procedure: A sample of 5g ± l mg was weighed into an extraction thimble and covered with

cotton that previously extracted with hexane (No.9-16-24/25-29-51, LOBA Cheme, and India).

Then, the sample and a pre-dried and weighed extraction flask containing about 100 ml hexanes

were attached to the extraction unit (Electro thermal, England) and the extraction process was

conducted for 6 hrs. At the end of the extraction period, the flask was disconnected from the unit

and the solvent was redistilled. Later, the flask with the remaining crude ether extract was put in

an oven at 105 °C for 3 hrs, cooled to room temperature in a desiccator, reweighed and the dried

extract was registered as fat content according to the following formula: Fat content (%) = (W2-

W1) × 100 % W3 Where; W2 =Weight of the flask and ether extract W1 =Weight of the empty

flask W3=initial weight of the sample 21.

2.11.1.4 ASH CONTENT

The ash content was determined according to the method described by the AOAC (2003).

Principle: The inorganic materials which are varying in concentration and composition are

customary determined as a residue after being ignited at a specified heat degree. Procedure: A

sample of 5g ±1 mg was weighed into a pre-heated, cooled, weighed and tarred porcelain

crucible and placed into a Muffle furnace (No.20. 301870, Carbolite, England) at 550 to 600 °C

until a white gray ash was obtained. The crucible was transferred to a desiccator, allowed to cool

to room temperature and weighed. After that, the ash content was calculated as a percentage

based on the initial weight of the sample. Ash (%) = [(Wt of crucible +Ash) - (Wt of empty

crucible)] x 100% Initial weight (Wt)

33
2.11.1.5. TOTAL CARBOHYDRATES

Total carbohydrates were calculated by difference according to the following equation:

Total carbohydrates = 100% - (Moisture + Protein + Fat + Ash).

34
 CHAPTER THREE

RESULT

Total Heterotrophic Bacteria count of different Plantain samples

Table 4.1: NA: Total Heterotrophic Bacteria count of different Plantain samples

S/No Sample Code Dilutions Cfu/g Log10

1 RP1 10-1 5.2 x103 3.716
2 RP2 10-1 5.4*103 3.73
3 RP3 10-1 5.9*103 3.77
4 RP4 10-2 6x103 3.78
5 RP5 10-2 6.2 x103 3.79
6 UP1 10-1 9.3 x103 3.97
7 UP2 10-1 0 0
8 UP3 10-2 0 0
9 UP4 10-2 9.7x103 3.99
10 UP5 10-2 0 0
11 RRP1 10-1 1.3 x103 3.11
12 RRP2 10-1 2.7 x104 4.43
13 RRP3 10-1 4.8 x103 3.68
14 RRP4 10-2 2.2x103 3.34
15 RRP5 10-2 4.7x104 4.67
16 URP1 10-1 5,2 x103 3.72
17 URP2 10-1 4.8 x103 3.68
18 URP3 10-1 3.62 x104 4.56
19 URP4 10-2 4.1 x104 4.61
20 URP5 10-2 6.55 x103 3.82
21 RBP1 10-1 0 0
22 RBP2 10-1 0 0
23 RBP3 10-1 3.26 x104 4.51

35
24 RBP4 10-2 1.04 x104 4.02
25 RBP5 10-2 0 0
26 UBP1 10-1 0 0
27 UBP2 10-1 0 0
28 UBP3 10-1 0 0
29 UBP4 10-2 0 0
30 UBP5 10-2 0 0
KEYSRP= Ripe Plantain

UR= Unripe Plantain

RRP= Ripe Roasted Plantain

URP= Unripe Roasted Plantain

RBP= Ripe Boiled Plantain

UBP= Unripe Boiled Plantain

36
Table 4.2: MSA: Total Heterotrophic Staphylococcus count of different Plantain samples

S/No Sample Code Dilutions Cfu/g Log10

1 RP1 10-1 2.4x103 3.38
2 RP2 10-1 1.47 x104 4.17
3 RP3 10-1 0 0
4 RP4 10-2 3.2 x104 4.51
5 RP5 10-2 3.9 x103 3.59
6 UP1 10-2 0 0
7 UP2 10-2 0 0
8 UP3 10-4 0 0
9 UP4 10-2 0 0
10 UP5 10-2 0 0
11 RRP1 10-4 0 0
12 RRP2 10-4 0 0
13 RRP3 10-3 0 0
14 RRP4 10-3 0 0
15 RRP5 10-4 0 0
16 URP1 10-2 0 0
17 URP2 10-3 0 0
18 URP3 10-2 0 0
19 URP4 10-3 0 0
20 URP5 10-3 0 0
21 RBP1 10-6 0 0
22 RBP2 10-3 0 0
23 RBP3 10-3 0 0
24 RBP4 10-3 0 0
25 RBP5 10-3 0 0
26 UBP1 10-2 0 0
27 UBP2 10-2 0 0
28 UBP3 10-4 0 0

37
29 UBP4 10-3 0 0
30 UBP5 10-2 0 0

KEYS

RP= Ripe Plantain

UR= Unripe Plantain

RRP= Ripe Roasted Plantain

URP= Unripe Roasted Plantain

RBP= Ripe Boiled Plantain

UBP= Unripe Boiled Plantain

38
Table 4.3: MAC:Total Coliform count of different Plantain samples

S/No Sample Code Dilutions Cfu/g Log10

1 RP1 10-1 0 0
2 RP2 10-1 0 0
3 RP3 10-1 0 0
4 RP4 10-2 0 0
5 RP5 10-2 0 0
6 UP1 10-1 0 0
7 UP2 10-1 0 0
8 UP3 10-2 0 0
9 UP4 10-2 0 0
10 UP5 10-2 0 0
11 RRP1 10-1 4.3×103 3.63
12 RRP2 10-1 0 0
13 RRP3 10-1 0 0
14 RRP4 10-2 0 0
15 RRP5 10-2 0 0
16 URP1 10-1 4.6x103 3.66
17 URP2 10-1 0 0
18 URP3 10-1 0 0
19 URP4 10-2 0 0
20 URP5 10-2 0 0
21 RBP1 10-2 0 0
22 RBP2 10-3 0 0
23 RBP3 10-3 0 0
24 RBP4 10-3 0 0
25 RBP5 10-3 0 0
26 UBP1 10-2 3x103 3.48
27 UBP2 10-2 0 0
28 UBP3 10-4 0 0

39
29 UBP4 10-3 0 0
30 UBP5 10-2 0 0

KEYS

RP= Ripe Plantain

UR= Unripe Plantain

RRP= Ripe Roasted Plantain

URP= Unripe Roasted Plantain

RBP= Ripe Boiled Plantain

UBP= Unripe Boiled Plantain

40
Table 4.3: SDA: Total Fungi count of different Plantain samples

S/No Sample Code Dilutions Cfu/g Log10

41
1 RP1 10-1 4.0 x104 4.6
2 RP2 10-1 1.47 x103 3.16
3 RP3 10-1 3.3 x103 3.51
4 RP4 10-2 0 0
5 RP5 10-2 5.0 x103 3.69
6 UP1 10-1 2.4 x104 4.38
7 UP2 10-1 4.2 x104 4.62
8 UP3 10-2 2.5 x104 6.39
9 UP4 10-2 0 0
10 UP5 10-1 9.7 x104 4.98
11 RRP1 10-1 3.3 x103 3.51
12 RRP2 10-2 5.0 x103 3.69
13 RRP3 10-1 0 0
14 RRP4 10-2 1.28 x103 3.10
15 RRP5 10-2 2.4 x104 4.38
16 URP1 10-2 4.6 x104 4.63
17 URP2 10-1 2.5 x104 4.39
18 URP3 10-1 0 0
19 URP4 10-1 5.1 x104 4.70
20 URP5 10-2 6.55 x104 4.81
21 RBP1 10-2 0 0
22 RBP2 10-2 0 0
23 RBP3 10-1 0 0
24 RBP4 10-1 0 0
25 RBP5 10-1 0 0
26 UBP1 10-2 0 0
27 UBP2 10-2 0 0
28 UBP3 10-4 0 0
29 UBP4 10-3 0 0
30 UBP5 10-2 0 0

42
KEYS

RP= Ripe Plantain

UR= Unripe Plantain

RRP= Ripe Roasted Plantain

URP= Unripe Roasted Plantain

RBP= Ripe Boiled Plantain

UBP= Unripe Boiled Plantain

Table1. pH values of plantain

Samples code RP UP RRP URP RBP UBP

43
pH 5.87 4.94 5.37 5.28 5.05 7.67

KEYS

RP= Ripe Plantain

UR= Unripe Plantain

RRP= Ripe Roasted Plantain

URP= Unripe Roasted Plantain

RBP= Ripe Boiled Plantain

UBP= Unripe Boiled Plantain

4.5
4.078
4 3.846
3.7572

3.5

2.5

2
1.706
1.592
1.5

0.5

0
0
RP UP RRP URP RBP UBP

Figure 4.4: Total Mean of Total Heterotrophic Bacteria Count of the different Plantain
samples

44
KEYS

RP= Ripe Plantain (3.7572)

UR= Unripe Plantain (1.592)

RRP= Ripe Roasted Plantain (3.846)

URP= Unripe Roasted Plantain (4.078)

RBP= Ripe Boiled Plantain (1.706)

UBP= Unripe Boiled Plantain (0)

3.5

2.5

1.5

0.5

0
RP UP RRP URP RBP UBP

Figure 4.8: Total Mean of Total Staphylococcus Count of the different Plantain samples

KEYS

RP= Ripe Plantain (3.3)

45
UR= Unripe Plantain (0)

RRP= Ripe Roasted Plantain (0)

URP= Unripe Roasted Plantain (0)

RBP= Ripe Boiled Plantain (0)

UBP= Unripe Boiled Plantain (0)

46
 0.8

0.7

0.6

0.5

0.4

0.3

0.2

0.1

0
RP UP RRP URP RBP UBP

Figure 4.12: Total Mean of Total Coliform Count of the different Plantain samples

KEYS

RP= Ripe Plantain (0)

UR= Unripe Plantain (0)

RRP= Ripe Roasted Plantain (0.726)

URP= Unripe Roasted Plantain (0.732)

RBP= Ripe Boiled Plantain (0)

UBP= Unripe Boiled Plantain (0.696)

47
TABLE 4.4: CHARACTERISTICS OF BACTERIA ISOLATED FROM RIPE AND UNRIPE PLANTAIN SAMPLES

ISOLATE COLOR SIZE SHAPE SURFACE MARGIN ELEVA- OPACITY CONSIS-

CODE TION TENCY
RP1 Milky 3mm Round Smooth/Shiny Undulate Flat Opaque Smooth
RP2 Yellow 2mm Round Dull Entire Convex Opaque Smooth
RP3 Cream 3mm Round Shiny Entire Raised Trans- Smooth
parent

RP4 Milky 4mm Spindle Shinny Curled Flat Opaque Mucoid

RP5 White 3mm Irreg- Dull Lobate Flat Opaque Mucoid
ular

UP6 Cream 1mm Round Shiny Entire Raised Trans- Smooth

parent
UP7 White 3mm Filametous Dull Lobate Flat Opaque Mucoid
UP8 Milky 2mm Round Dull Entire Convex Opaque Smooth
UP9 Yellow 1mm Filamentous Dull Undulate Convex Opaque Smooth
UP10 Yellow 4mm Round Shiny Entire Flat Opaque Smooth

48
TABLE 4.5: CHARACTERISTICS OF BACTERIA ISOLATED FROM ROASTED RIPE AND UNRIPE PLANTAIN
SAMPLES
ISOLATE COLOR SIZE SHAPE SURFACE MARGIN ELEVA- OPACITY CONSIS-
CODE TION TENCY
RRP1 Milky 3mm Round Smooth Lobate Flat Opaque Mucoid
RRP2 Yellow 3mm Round Dull Entire Convex Opaque Smooth
RRP3 Cream 4mm Round Shiny Curled Raised Trans- Smooth
Parent

RRP4 Milky 3mm Irreg- Dry Lobate Flat Opaque Mucoid

ular
RRP5 Opaque 2mm Filamentous Shiny Entire Raised Opaque Smooth
URP6 Cream 3mm Round Shiny Entire Raised Trans- Smooth
Parent
URP7 Milky 3mm Irreg- Dull Irreg- Flat Opaque Smooth
ular Ular
URP8 Milky 2mm Round Shiny Entire Convex Opaque Smooth

URP9 Yellow 2mm Round Dull Lobate Erose Opaque Mucoid

URP10 Cream 4mm Round Shiny Entire Raised Trans- Smooth
Parent

49
TABLE 4.6: CHARACTERISTICS OF BACTERIA ISOLATED FROM BOILED RIPE AND UNRIPE PLANTAIN
SAMPLES

ISOLATE COLOR SIZE SHAPE SURFACE MARGIN ELEVA- OPACITY CONSIS-

CODE TION TENCY
RBP1 Yellow 2mm Irreg- Shinny Curled Flat Opaque Smooth
ular
RBP2 Yellow 2mm Round Dry Entire Flat Opaque Smooth
RBP3 Milky 3mm Round Shiny Entire Flat Opaque Smooth
RBP4 Cream 1mm Round Smooth Curled Raised Trans- Smooth
parent
RBP5 Milky 2mm Round Shiny Entire Flat Opaque Smooth
UBP6 Yellow 3mm Round Shiny Curled Flat Opaque Smooth
UBP7 Cream 2mm Round Shiny Entire Raised Opaque Smooth
UBP8 Cream 3mm Round Shiny Curled Raised Opaque Smooth
UBP9 White 2mm Irreg- Smooth Irreg- Flat Opaque Smooth
ular ular
UBP10 Cream 1mm Round Shiny Entire Raised Opaque Smooth

50
TABLE 4.7: BIOCHEMICAL CHARACTERISTICS OF BACTERIA ISOLATED FROM RIPE AND UNRIPE PLANTAIN
SAMPLES

Cell Morphology
Gram Reaction

Glucose

Sucrose
Isolate code

Lactose
Catalase

Oxidase

Motility
Citrate

Indole

Slant
Butt

Gas
MR

H2S
VP
A G A G A G
RP1 + Rod + + - + + - + - - + + - - B - B - Bacillus subtilis
RP2 + Rod + + - - + + - - - - - - - B - B - Bacillus subtilis
RP3 + Cocci + + - - - + + + - + - + - B - B - Staphylococcus sp
RP4 + Rod + - + - + - + - - - - - - B - B - Bacillus subtilis
RP5 - Rod + + + + + + - - - + + - - A - B + Proteus sp
UP6 + Cocci + + - - + + + - - - - - - B - B - Staphylococcus sp
UP7 - Rod + + - - + - - - - + + - - B - B - Proteus
UP8 + Rod + + - - - - + - - + + - - A + B + Bacillus subtilis
UP9 + Cocci + + + - + - - - - - - - - B - B - Streptococcus sp
UP1 + Rod + + + - + - - - - + - + - A - B - Bacillus subtilis
0

Keys: + = Positive, - = Negative, A = Acid, B = Base, A &G = Acid and Gas

51
 TABLE 4.8: BIOCHEMICAL CHARACTERISTICS OF BACTERIA ISOLATED FROM ROASTED RIPE AND UNRIPE
PLANTAIN SAMPLES

Gram Reaction

Morphology

Glucose

Sucrose
Isolate code

Lactose
Catalase

Oxidase

Motility
Indole

Slant
Butt
Cell

Gas
MR

H2S
VP
A G A G A G
RRP1 + Rod + + - + - + - - + + + + B + B + Bacillus subtilis
RRP2 + Cocci + + - + - + - - + + - - A + B + Micrococcus sp
RRP3 + Cocci + - - - + + + + + + + + B - B - Staphylococcus sp
RRP4 + Rod + + - + - + - - - - - - B - B - Bacillus subtilis
RRP5 + Cocci + - - + + - - - + + - + A + A + Proteus sp
URP6 + Rod + - - - + + + + + + + - A - A - Staphylococcus sp
URP7 + Cocci + - - - + + + - + + + - B - A + Micrococcus sp
URP8 + Rod + - - + - + - - + - - + B - B + Bacillus cereus
URP9 + Cocci + + - + + + + + + + - - B + B + Staphylococcus sp
URP10 - Cocci + - - - - - + + + + + + B + B + Staphylococcus sp

Keys: + = Positive , - = Negative, A = Acid, B = Base, A &G = Acid and Gas

52
TABLE 4.9.: BIOCHEMICAL CHARACTERISTICS OF BACTERIA ISOLATED FROM BOILED RIPE AND UNRIPE
PLANTAIN SAMPLES

Gram Reaction

Morphology

Glucose

Sucrose
Isolate code

Lactose
Catalase

Oxidase

Motility
Citrate

Indole

Slant
Butt
Cell

Gas
MR

H2S
VP
A GA G A G
RBP1 + Rod + + + - + - + - - - - - - B - B - Bacillus subtilis
RBP2 + Rod + + - - + - + - - - - - - B - B - Bacillus subtilis
RBP3 + Cocci + - + - + - + - - - - - - B - B - Staphylococcus sp
RBP4 + Rod + + + - + - + - - - - - - B - B - Bacillus sp
RBP5 + Cocci + + - - + - + - - - - - - B - B - Staphylococcus sp
UBP6 + Rod + + + - + + - - - + + - - B - B + Bacillus sp
UBP7 + Rod + + + - + + - - - + + - - B - B - Bacillus cereus
UBP8 + Cocci + - - - + - - - - - - + + B - B - Staphylococcus sp
UBP9 + Rod + + - - + - - - - - - + + B - B - Bacillus cereus
UBP1 + Rod + + + - + + - - - + + - - B - B - Bacillus cereus
0

Keys: + = Positive , - = Negative, A = Acid, B = Base, A &G = Acid and Gas

53
 Streptococcus
10%

Proteus sp
20%

Bacillus sp
50%

Staphy-
lococcus
sp
20%

Bacillus sp Staphylococcus sp Proteus sp Streptococcus

Fig. 4.13: Percentage frequency of Bacteria Isolated from Ripe n unripe Plantain

KEYS

Bacillus sp

Staphylococcus sp

Proteus sp

Streptococcus sp

54
 Micro-
coccus
20%
Bacillus sp
30%

Proteus sp
10%

Staphylococcus sp
40%

Bacillus sp Staphylococcus sp Proteus sp Micrococcus

Fig. 4.14: Percentage frequency of Bacteria Isolated from Roasted ripe n unripe

KEYS

Bacillus sp

Staphylococcus sp

Y
Proteus sp

Micrococcus sp

55
Fig. 4.15: Percentage frequency of Bacteria Isolated from boiled plantain samples

KEYS

Bacillus sp

Staphylococcus sp
Thamnid- As-
ium pergillus
14% 14%

Rhizopus
sp
7%
yeast
14%
Mucor
7%

Penicillum Cladospo-
14% rium
14%

Fusarium
14%

Aspergillus yeast Cladosporium Fusarium Geotrichum

Penicillum Mucor Rhizopus sp Thamnidium

Fig. 4.16: Percentage frequency of Fungi Isolated from all Ripe plantain samples

56
 Thamnid- As-
ium pergillus
14% 14%

Rhizopus
sp
7%
yeast
14%
Mucor
7%

Penicil- Cladospo-
lum rium
14% 14%

Fusarium
14%

Aspergillus yeast Cladosporium Fusarium Geotrichum

Penicillum Mucor Rhizopus sp Thamnidium

Fig. 4.17: Percentage frequency of Fungi Isolated from all unripe plantain samples

57
TABLE 10: CHARACTERISTICS AND IDENTIFICATION OF FUNGI ISOLATE

Microscopic Examination MicrobialSp

IsolateCode Characteristics Morphology
Candiospores appears smooth
double storigmata, seen
Colonies appear black, black with withspot on vesicle condidia Aspergillus
fast growing mycelium with somehow spherical sp.
RP1 spores
Irregular branched conidia
sphores bearing a whool of
Fluffy spread with a diameter of phialide conidia hyaline Fusarium sp
UP2 7mm in three Days, creating white
Non- septate hyphae, has
sporangio sphore white bears a Rhizopus
terminal sporangium sp
RRP3 Light brown mycelium
Condiospores long, slender
septate above philiades bear Penicillium
Colonies appear grayish green single condia hyaline sp
RUP4 which turns white.
Aseptate hyphae, long
branched sporangiophore
Colonies are brown and Mucoid in bearing sporangium Mucos sp
RRP4 appearance
Beige in color, 0.075mm 75 mm
in diameter sphericalo egg-shaped Oval (egg shaped) organism Yeast sp
RP6 to filamentous
Filamentous, with velvety
white mycelia on the surface of Thamnidium
45 mm diameter at 28 •C, 1-4mm colony. spp
UP7 high, flat, with white velvety
RBP No Growth
UBP No Growth

58
 MICROBIAL GROUP RIPE PLANTAIN RIPE ROASTED RIPE BOILED
PLANTAIN PLANTAIN

FUNGI ISOLATE FROM RIPE VARIOUS SAMPLES

Aspergillus sp Present Present Absent

Yeast sp Presnt Present Absent

Cladosporium sp Present Present Absent

Fusarium sp Present Present Absent

Geotrichum sp Absent Absent Absent

Penicillum sp Present Present Absent

Mucor sp Present Absent Absent

Rhizopus sp Present Absent Absent

Thamnidium Present Present Absent

59
 MICROBIAL GROUP UNRIPE PLANTAIN UNRIPE ROASTED UNRIPE BOILED
PLANTAIN PLANTAIN

FUNGI ISOLATE FROM UNRIPE VARIOUS SAMPLES

Aspergillus sp Present Present Absent

Yeast sp Presnt Present Absent

Cladosporium sp Present Present Absent

Fusarium sp Present Present Absent

Geotrichum sp Absent Absent Absent

Penicillum sp Present Present Absent

Mucor sp Present Absent Absent

Rhizopus sp Present Absent Absent

Thamnidium Present Present Absent

60
PROXIMATE COMPOSITION RESULTS

The proximate composition of the samples is shown in Table 1 while Table 2 shows the results
of the mineral analysis of the samples.
Table 1: Proximate Analysis of the Samples
Amount (%)
S/N Parameters Ripe Plantain Pulp Unripe Plantain Pulp

1 Moisture Content 58.24 36.66

2 Ash Content 3.10 1.20

3 Fat Content 1.30 6.69

4 Crude Fiber 2.89 3.53

5 Protein Content 3.50 5.14

6 Carbohydrate Content 30.33 47.42

61
Table 2: Mineral Analysis of the Samples
Concentration (mg/kg)
S/N Mineral Ripe Plantain Pulp Unripe Plantain Pulp
1 Fe 12.40 33.45

2 Ca 141.60 132.40

3 Mg 34.25 34.30

4 Mn 0.50 2.20

5 Zn 4.65 13.35

6 Cu Nil Nil

62
 CHAPTER FOUR

4.0 DISCUSSION AND CONCLUSION

The data presented shows the microbial quality and sensory characteristics of ripe, unripe

plantain, ripe and unripe roasted plantain, ripe and unripe boiled plantain obtained from different

locations in Rivers State, Port Harcourt. The pattern of distribution showed that most of the

isolates found in ripe and unripe plantain were obtained from hawkers and market women.

Samples investigated harbored a wide variety of organisms. Fruits with mechanical injuries from

the farm, as a result of transportation or handling were also examined for the type of organisms

that entered the wounds thereby spoiling the fruits. Bacterial and fungi can be sucked into

plantain plant through natural plant openings such as stomata, hydathodes or lenticels. They can

enter through abrasions or wounds on leaves, stems or roots or through placement by specific

feeding insects.

The total heterotrophic count counts (TVC) ranged from 5.4 - 6.2 x 10 3 Cfu/g in raw ripe

plantains, 0 – 9.7 x 103, 1.3 x 103 – 4.7 x 104 Cfu/g in ripe roasted, 4.8 x 10 3 – 4.1 x 104 Cfu/g in

unripe roasted and 1.04 – 3.26 x104- in ripe boiled while unripe boiled plantains gave no counts.

This is high in comparison with the work of (REFERENCE). The total Staphylococcus count

counts ranged from 2.4 x 103- 3.2 x 104 Cfu/g in raw ripe plantains, while all other plantain

63
samples gave no counts. The total coliform count (TCC) counts ranged from 0 - 4.3 x 10 3 Cfu/g

in ripe roasted plantains, 0 – 4.6 x 103 Cfu/g in unripe roasted plantain and 0 -3 x 10 3 Cfu/g for

boil unripe plantain while all other plantain samples gave no counts. The toto fungal counts

(TFC) ranged from 0 - 4.0 x 104 Cfu/g in raw ripe plantains, 0 – 9.7 x 10 4 in unripe raw, 0 x – 2.4

x 104 Cfu/g in ripe roasted, 0 – 6.55 x 104 Cfu/g in unripe roasted plantains. All other samples

gave no fungal counts.

Most of the organisms encountered could be found in soil, dust and on the bodies of insect,

animals and humans (Fagbohun et al., 2010). Staphylococcus aureus is an indicator of

contaminations from human sources. It is a normal flora of the skin of man and could have been

into the product during roasting of plantain (Fagbohun et al., 2010) staphylococcus aureus is

known to produce enterotoxin leading to food intoxication. Bacillus subtilis is an indicator of

contamination from environmental sources; it produces endospores which can help it to survive

the heat treatment and is associated with miscellaneous problems. pseudomonas aeruginosa is

also an indicator of possible post- production contamination. These suggest that the samples

could serve as common vehicle for pathogens transmitted via the feacal oral route. The presence

of Aspergillus Niger, Rhizopus stolonfer, Mucor mucedo, yeast sp, thamnidium sp and

Penicillium sp. on these products buttress the likelihood of post-production contamination.

Some of the fungi, most especially Aspergillus sp. Some strains of Aspergillus niger have been

reported to produce potent mycotoxins called ochratoxins. Penicillium infections results in

keratitis, endophtalmitis, otomycosis, necrotizing esophagitis, pneumonia, endocarditis,

peritonitis, urinary tract infections, mucocutaneous, genitourinary, gastrointestinal, pulmonary

and disseminated infections as the clinical features (Kontogiorgi et al., 2007).

64
Aspergillus species are filamentous fungi that are commonly found in soil, decaying vegetation,

and seeds and grains, where they thrive as saprophytes. Aspergillus species can be occasionally

harmful to humans. Most Aspergillus species are found in a wide variety of environments and

substrates on the Earth throughout the year. Only a few well-known species are considered as

important opportunistic pathogens in humans.

In humans, Aspergillus fumigatus is the most common and life-threatening airborne opportunistic

fungal pathogen, which is particularly important among immunocompromised hosts.

There is considerable concern regarding the potential health outcomes of exposure to biological

materials existing in the air. Molds constitute an important threat to human health; their effects

range from moderate allergies and severe asthma to disseminated infections. Exposure to molds

in indoor places is not typically considered a specific risk factor in the etiology of fungal diseases

unless some special conditions are present that are essential for specific infections.

Fungal infections that are particularly aggressive to tissues are limited to immunocompromised

individuals (e.g., hospitalized patients). Aspergillus species are a ubiquitous mold in home and

hospital environments. Indoor plants represent a natural environment for the growth of these

fungi; however, few recommendations have been made about avoiding known sources of fungal

proliferation (plants and flowers) in indoor places.

Molds such as Aspergillus may adversely affect human health based on toxicity, allergy, and

infection. Some species of Aspergillus are known to be capable of producing secondary

metabolites or mycotoxins. Inhaling high concentrations of mixed organic dusts, including

65
mycotoxins, volatile organic compounds (VOCs), and allergens (glucans), are associated with

sick building syndrome.

The most important medical relevance of Fusarium spp. is based on their phytopathogenic

property, contributing to hunger and undernutrition in the world. A few Fusarium spp., such as

F. oxysporum and F. solani, are opportunistic pathogens and can induce local infections, i.e., of

nails, skin, eye, and nasal sinuses, as well as occasionally, severe, systemic infections, especially

in immunocompromised patients. These clinical diseases are rather difficult to cure by

antimycotics, whereby the azoles, such as voriconazole, and liposomal amphotericin B give

relatively the best results. There are at least two sources of infection, namely the environment

and the gut mycobiome of a patient. A marked impact on human health has the ability of some

Fusarium spp. to produce several mycotoxins, for example, the highly active trichothecenes.

These mycotoxins may act either as pathogenicity factors, which means that they damage the

host and hamper its defense, or as virulence factors, enhancing the aggressiveness of the fungi.

Acute intoxications are rare, but chronic exposition by food items is a definite health risk,

although in an individual case, it remains difficult to describe the role of mycotoxins for inducing

disease. Mycotoxins taken up either by food or produced in the gut may possibly induce an

imbalance of the intestinal microbiome. A particular aspect is the utilization of F. venetatum to

produce cholesterol-free, protein-rich food items.

None of the samples were visibly moldy which suggests that fungi were yet to be established;

hence, contamination is likely due to fungal propagules in the air. As a result of unhygienic

processing and packaging conditions, roasted plantain may be contaminated with

microorganisms during human and surface contact. It could also be as a result of environmental

66
exposure; improper storage of plantain fruits and insect contact that result in microorganisms’

contaminating plantain that are to be sold in the market.

When preparing boiled plantain, good production procedures and proper hygiene should be

observed to avoid contamination by bacteria and fungi. The number and type of microbes present

on the product are major determinant of quality deterioration. The products were found to

contain bacteria and fungi. These microbes multiply and lead to spoilage of the products and

render them inedible, making them a vehicle for food poisoning. Producers of these products

should also handle them in such a way that they prevent contamination by proper preparation and

proper packaging. In the case of roasted plantain (locally known as “bole”), packaging should be

used to avoid moisture from the atmosphere accumulating which might lead to deterioration of

the food quality. Various group of bacterial species obtained from these sources including L.

fermentum can actively utilize sugar in the plantain for their survival and Bacillus cereus is a

spoilage organism responsible for the spoilage of the plantain flour while the presence of the

other species of Bacillus is due to the presence of proteins and versatility of Bacillus species to

thrive well in diversified environment or form part of natural micro flora of plantain (Lebedaet

al.,2001). It was observed that there was high percentage of protein in ripe plantain pulp, Fe and

other minerals except cu were present in higher quantities in peels of ripe and unripe plantain

compared with that observed in the pulp. Therefore, it could be recommended that the peel

should be ground into fine powder and use it to fortify plantain pulp flour to increase its

nutritional value. The moisture content, Ash content and Crude fiber of ripe plantain have a

higher percentage than unripe plantain. While the fat content, protein content and carbohydrate

content of unripe plantain have a higher percentage than ripe plantain.

Proximate Analysis

67
The proximate parameters (moisture, ash, crude fat, crude fibre, crude protein and carbohydrate)

contents were determined using the procedure. The moisture content was determined using an

AND-MF-50 moisture analyzer at 105oC. The moisture content in each of the samples was

determined by weighing 3g of each sample into a pre-weighed aluminum dry dish, the samples

were dried to a constant weight in the moisture analyzer at 105oC for about 45 minutes and then

the moisture was read and recorded. The ash content was determined for all the samples by the

incineration of the samples placed in a muffle furnace maintained at 550oC for 5 – 8 hours while

the crude fibre obtained by digesting 2g of the samples with H2SO4 and NaOH and incineration

the residue in a muffle furnace maintain at 550oC for 5 – 8 hours. The crude protein (% total

nitrogen x 6.25) was determined by kjeldahl method, using 2g of each sample. The crude lipid

content was also determined by exhaustively extracting 10g of each sample in a soxhlet

apparatus using n-hexane as the extracting solvent and the carbohydrate content was determined

by deducting the total percentage of moisture, ash, fibre, fat and protein from 100.

Mineral Analysis

The mineral analysis was carried out using standard methods. The samples were dry ashing at

550oC to constant weight and dissolved the ash in volumetric flasks with de-ionized water with a

few drops of concentrated HCl. The sample solutions were stored for mineral analysis using

Atomic Absorption Spectrophotometer.

Moisture content

The moisture level of ripe and unripe plantain samples varied between 58.24% - 33.6%. Unripe

plantain has a significantly lower amount of moisture being 33.6%.

Ash content

68
The ash content of ripe plantain samples varied between 3.10% - 1.20%. Unripe plantain had the

lesser ash content which is the value of 1.2%.

Fat content

The lipid content level of ripe plantain samples varied between 1.30% -6.69%. Unripe plantain

had a higher fat content although not by a great margin.

Fibre content

The fibre content of ripe plantain samples varied between 2.89% -3.53%. The crude fibre in the

ripe plantain samples shows depreciation in the fruit while unripe plantain has about a 1%

difference higher in fiber content.

Protein content

The protein content of ripe plantain samples varied between 3.50% -5.14%. As expected, the

protein content in unripe plantain was found to be slightly higher than ripened plantain (5.14%).

Carbohydrate content

The carbohydrate content of ripe plantain samples varied between 30.33% -47.42%.

Carbohydrates were within range of 31% which is the standard for plantains. Unripe plantain

was significantly higher for these samples.

Plantain Nutritional Facts

137g of plantain provides 166 calories, 1.5g of protein, 40g of carbohydrates, and 0.1g of fat.

Plantains are an excellent source of vitamin C, fiber, and vitamin B6. The following nutrition

69
information is provided by the U.S. Department of Agriculture (USDA).

Calories: 166

Fat: 0.1g

Sodium: 2.7mg

Carbohydrates: 40g

Fiber: 3.5g

Sugars: 3.1g

Protein: 1.5g

Vitamin C: 12.5mg

Vitamin B6: 0.3mg

Plantains provide a healthy dose of carbohydrates. 137g green (unripe) plantains has 40 total

grams of carbohydrates, with nearly 4 grams of fiber and just 3 grams of natural sugar. As

plantains ripen, fiber content goes down and sugar content increases. Plantains are not a

significant source of protein. A medium plantain has less than 2 grams.

Vitamins and Minerals

Plantains contain iron, vitamin C, vitamin B6, folate, potassium, magnesium, copper, and

vitamin A.3 According to the USDA, a cup of plantains provides 12.5 milligrams of vitamin C,

which is about 15% of your daily recommended intake. Plantains contain folate, which is a vital

nutrient for women trying to conceive. You'll get nearly 20% of your daily recommended intake

from a cup of cooked plantains.

70
Calories

137g of unripe plantain provides 166 calories, 96% of which come from carbs, 3% from protein,

and 1% from fat.

4.1 FOOD SAFETY

According to the Centers for Disease Control and Prevention, in the U.S. the number of reported

produce-related outbreaks per year doubled between the period 1973-1987 and 1988-1992

(2,24). During both time periods, the etiologic agent was unknown in more than 50% of

outbreaks. (Beuchat, 2002). Escherichia coli O157:H7 infection has been associated with lettuce,

sprouts, and apple juice, and enterotoxigenic E. coli has been linked to carrots. Documented

associations of shigellosis with lettuce, scallions, and parsley; cholera with strawberries; parasitic

diseases with raspberries, basil, and apple cider; hepatitis A virus with lettuce, raspberries, and

frozen strawberries; and Norwalk/Norwalk-like virus with melon, salad, and celery have been

made. Among the greatest concerns with human pathogens on fresh fruits and vegetables are

enteric pathogens (e.g., E. coli O157:H7 and Salmonella) that have the potential for growth prior

to consumption or have a low infectious dose. Bacterial pathogens have been isolated from a

wide variety of fresh produce. (Beuchat, 2002).

4.2 RECOMMENDATION

This paper recommends that every vendor, helper or food handler should undergo a basic

training in food hygiene. This ensures that they follow the required rules for hygiene and

sanitation.

71
The government should invest in street food industries as it provides employment, cheap food,

and wide variety of food for the urban dwellers. Through the ministry of health and local

government, legislation should be developed to recognize the street foods industries by

developing code of practice for street food vending.

1) Provide clean prepared places for supply and selling of plantain fruit.

2) Consumers should avoid consumption of over ripe banana which may be contaminated with

microbes. According to the obtained results, we recommend discarding over ripened plantain

because of the presence of pathogenic microorganisms.

3) Further research is needed at different levels of production.

Plantains are of great economic importance, and in order to maintain quality, they should be

stored under proper-controlled conditions to prevent them from fungal deterioration. The present

study shows the nature of bacterial and fungal species associated with plantain. This can enhance

the quality for processing this crop for human consumption. The presence of various species of

fungi in this crop can result in the synthesis of toxins such as aflatoxins, fumonisins, and ergot

alkaloids, thereby posing threat to immunocompromised individual. Hence, proper storage of

plantain against contamination to meet the international standards of good manufacturing process

will facilitate sustainable economic growth and food safety.

REFERENCES

72
Adeniji, T.A., Barimalaa, I.S. & Achinewhu, S.C. (2006). Evaluation of bunch characteristics
and flour yield potential in black Sigatoka resistant plantain and
banana hybrids. Global Journal of Pure and Applied Science, 12, 41–43.

Anon. 1999. Two is enough – really. Sweet tidings on treating high blood pressure. Asiaweek.
March 19, 1999.

Berg, J. R., Berg, R. C. Sarna, E. J. & Bates, B. 1971. Banana and plantain products and process
for preparing same. British Patent1:232, 773.

Aguilera, M. O., Stagnitta, P. V., Micalizzi, B. and De Guzman, A. M. S. (2005). Prevalence and
characterization of Clostridium perfringens from spices in Argentina. Anaerobe, 11:327-
334.
Ahene, R. E., Odamtten, G. T. and Owusu, E. (2011). Fungal and bacterial contaminants of six
spices and spice products in Ghana. Afr J Environ Sci Technol. 5(9):633–40.
Alam-Khan, K. and Abrahem, M. (2010). Effect of irradiation on quality of spices. Int Food Res
J. 17:825–36.
Aziz, N. H., Youssef, Y. A., El-Fouly, M. Z. and Moussa, L. A. (1998). Contamination of
some common medicinal plant samples and spices by fungi and their mycotoxins.
Bot. Bull. Acad. Sci., 39: 279- 285.
Badiru, I. and Badiru, D. (2013). Isi Cookbook: Collection of Easy Nigerian Recipes.
Bloomington: iUniverse. p. 36.
Banarjee, M. and Sarkar, P. K. (2003). Microbiological quality of some retail spices in India.
Food. Res. Int. 36:469-474.
Beuchat, L., Komitopoulou, E., Betts, R., Beckers, H., Bourdichon, F. and Joosten, H. (2011).
Persistence and survival of pathogens in dry foods and dry food processing
environments. Belgium: International Life Science Institute, Europe. Pp. 164-175.
Beuchat, L. R. (2002). Ecological factors influencing survival and growth of human pathogens
on raw fruits and vegetables. Microbes Infect. 4:413-423.
Beuchat, L. R., Komitopoulou, E., Beckers, H., Betts, R. P., Bourdichon, F. and Fanning, S.
(2013). Low-water activity foods: increased concern as vehicles of foodborne pathogens.
J Food Prot. 76(1):150-172.
Buckenhüskes H. J. and Rendlen M. (2004). Hygienic problems of phytogenic raw
materials for food production with special emphasis to herbs and spices. Food Sci.
Biotechnol., 13: 262–268.
Burnett, S. L., Gehm, E. R., Weissinger, W. R. and Beuchat, L. R. (2000). Survival of
Salmonella in peanut butter and peanut butter spread. J Appl Microbiol. 89(3):472-7.

73
Bowrey, R.G., Buckle, K.A., Hamey, I. & Pavenayotin, P. (1980). Use of solar energy for
banana drying. Food Technology in Australia, 32, 290–291.

Biyani, Manish; Tamiya, Eiichi; Takamura, Yuzuru; Saito, Masato; Ushijima, Hiromi; Biyani,
Radhika; Biyani, Madhu (2018-04-05). "Instant enumeration of total viable bacterial
counts for food quality assurance using 'DEP-On-Go' sensor. Its scope is as an
assessment tool rather than focus towards a specific organism". Analytical Methods.
Royal Society of Chemistry. 10 (14): 1585–1592. "Labeling & Nutrition" Guidance for
Industry: A Food Labeling Guide (7. Nutrition Labeling; Questions G1 through P8)

Chandler S. 1995. The nutritional value of bananas.Pp; 486-480 in Bananas and Plantains (S.
Gowen, ed.). Chapman & Hall, UK.

Daniells, J., Jenny, C., Karamura, D., and Tomekpe,K. 2001. Musa of Musa germplasm.
Diversity in the genus Musa (E. Arnaud and S. Sharrock, compil.). International
Network for the Improvementof Banana and Plantain (INIBAP), Montpellier, France.
<www.inibap. org/publications/musalogue.pdf>.3-336
.
Demirel, D. & Turhan, M. (2003). Air drying behaviour of dwarf Cavendish and Gros
Michel Banana slices. Journal of Food Engineering, 59, 1–11.

Falade K.O. & Olugbuyi A.O. (2009). Effects of maturity and drying methods on the physico-
chemical and reconstitution properties of plantain flour. International Journal of Food
Science and Technology 2010, 45, 170–178.

Fagbohun, E. D., Abegunde O. K. and David, O. M. (2010).Nutritional and mycoflora changes

during storage of plantain chips and the health implications.Journal of Agricultural
Biotechnology and Sustainable Development, 2(4): 61-65.
Fajinmi, O.B., Akinyemi, S.O.S and Aduramigba-Modupe, A.O. (2011). Influence of
storage methods on ripening and microbial loads of some Musa species.
Agriculture and Biology of North America, 2011, 2(8): 1201-1207.

Kontogiorgi, M., Floros, I., Koroneos, A., Vamvonka, C., Paniara, O., Roussos, C., Routi,
C.b(2007).Fatal post-truamaticzygomycosis in an immunocompetent young patient.J. Medical
Microbiology, 56: 1243-1245.

Lebeda, A., Luhová, L., Sedlářová, M., Jančová, D. (2001).The role of enzymes in plant-fungal
pathogens interactions. Journal of Plant Diseases and Protection, 108: 89-111.

Li, S., Hartman, G.L. and Widholm, J.M. (1999). Viability staining of soybean
suspension-cultured cells and a seedling stem cutting assay to evaluate phytotoxicity of
Fusariumsolanif. sp. Glycinesculture filtrates. Plant Cell Report.18: 375-380.

74
Ohenhen, R.E., Enweani, L.B., Ogiehor, S.I. and Uwabor, K. (2013). Microorganisms
associated with preparation of plantain pudding in Western Nigeria. African Journal
of Microbiology, 1(2): 037 – 039.

Ruiz,O.H., Gómez,C.B., Colmenares, N.A.and Cooman,A. (2009). Guide Identification and

Integrated Pest Management in Banana and Plantain in Magdalena and Uraba
Colombia: “Reduction of pesticide runoff into the caribbean sea”. Edition and
Design: COMUNICACIONES AUGURA Printing: IMPRESOS S.A. Medellín–
Colombia.ISBN 978-958-99167-0-4. Pp. 64

75
 APPENDIX

PLATE COUNT AGAR

Formulation Gms/L

Enzymatic Digest of casein/tryptone 5.0

Yeast Extract 2.5
Glucose 1.0
Agar 15.0

PH = 7.4+/- 0.2 at 250c

SABROUD DEXTROSE AGAR

Formulation Gms/L
Balance peptone no1 10.0
Dextrose 40.0
Agar 15.0
PH = 7.4+/- 0.2 at 250c

MANNITOL SALT AGAR

Formation Gms/L
Pancreatic Digest of Animal Tissue 5.0
Peptic Digest of Animal Tissue 5.0
Beef Extract 1.0
Sodium chloride 75.0
D-mannitol 10.0
Phenol Red 0.025
Agar 15.0
PH = 7.4+/- 0.2 at 250c

76
MacConkey Agar

Formation Gms/L
Peptone (Pancreatic digest of gelatin) 17.0
Proteose (meat and casein) 3.0
Laclose monohydrate 10.0
Bile salts 1.50
Sodium chloride 5.0
Neutral Red 0.03
Crystal Violet 0.001
Agar 13.50
H 0
P = 7.4+/- 0.2 at 25 c

NORMAL SALINE
Nacl 8.5g

Distilled water 1000g

LACTOPHENOL

Phenol 10g

Cotton 0.04g

Lactic acid 10ml

Glycerol 20ml
-

PEPTONE WATER

Peptone 2g

77
Nacl 1g

Distilled water 1000ml

INDOLE

Tryptone 10g

Nacl 5g

Distilled water 1000ml

pH 7.2

78