MICROBIAL ANALYSIS OF FERMENTED AND UNFERMENTED SWEET POTATO-

PIGEON PEA WEANING FOOD

BY

OLAGBE BLESSING TOYIN

SO4/FST/2019/2375

A PROJECT SUBMITTED TO THE DEPARTMENT OF FOOD SCIENCE AND

TECHNOLOGY, FACULTY OF APPLIED SCIENCES, RUFUS GIWA
POLYTECHNIC, OWO, ONDO STATE, NIGERIA

IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE AWARD OF

NATIONAL DIPLOMA (ND) IN FOODSCIENCE AND TECHNOLOGY

MARCH, 2022.

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 CERTIFICATION

This is to certify that this project was carried out by Olagbe Blessing Toyin, in the
department of Food Science and Technology, Faculty of Applied Sciences, Rufus Giwa
Polytechnic, Owo, Ondo State. In partial fulfillment of the requirement for the Award of
National Diploma (ND)

Mrs. Orungbemi, O.O ………………………………......

Project Supervisor Signature & Date

Dr. (Mrs.) Agbaje, R.B ……………………………………

Head of Department Signature & Date

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 DEDICATION

This project is dedicated to the Glory of Almighty God and to my parent Mr. and Mrs.

Olagbe through which her moral, spiritual and financial assistance I was able to attain my

dream.

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 ACKNOWLEDGEMENTS

With gladness and joyous of heart, I give thanks and glory to Almighty God who made it

possible for me to witness the end of this project without any hindrance I can't do but also show

my appreciation to my HOD Dr. (Mrs.) Agbaje R.B. For being very supportive, their parental

care and love makes this project attain the greatest success.

I also say a big thanks to my supervisor Mrs. Orungbemi, O.O, who gives relentless effort

and being very supportive throughout my project activities.

Also my profound gratitude goes to my parents Mr. and Mrs. Olagbe, who gives relentless

support in terms of advice and financial support in the completion of my programme (National

Diploma), I pray that Almighty God continue to support them in all their endeavors

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 TABLE OF CONTENTS

Title page i

Certification ii

Dedication iii

Acknowledgements iv

Table of contents v

List of Tables viii

List of Figures ix

Abstract x

CHAPTER ONE

1.0 Introduction 1

1.1 Aim and Objective of the Study 3

CHAPTER TWO

2.0 LITERATURE REVIEW 4

2.1 Sweet Potato (Ipomoea batatas) 4

2.1.1 Production and Consumption 5

2.1.2 Classification and Origin 6

2.1.3 Postharvest Handling Practices 8

2.1.4 Nutritional Composition of Sweet potato 10

2.1.5 Processing and Utilization 13

2.1.6 Health Benefits of Sweet Potato 15

2.2 Pigeon Pea (Cajanus cajan (L) Millsp) 16

2.2.1 Origin of Pigeon Pea 17

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2.2.2 Botany 17

2.2.3 Nutritional composition of pigeon pea 18

2.2.4 Health benefits of Pigeon Pea 23

2.2.5 Use as functional foods 27

2.2.6 Socio-economic Impact of Pigeon Pea 28

CHAPTER THREE

3.0 Materials and Methods 30

3.1 Materials 30

3.2 Methods 30

3.2.1 Preparation of fermented sweet potato-pigeon pea flour 30

3.2.2 Preparation of unfermented sweet potato-pigeon pea flour 30

3.3 Microbial Evaluation of weaning food 35

CHAPTER FOUR

4.0 Results and Discussion 38

4.1 Results 38

4.2 Discussion 39

CHAPTER FIVE

5.0 Conclusion and Recommendations 41

5.1 Conclusion 41

5.2 Recommendations 41

REFERENCES 42

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 LIST OF TABLES

Table Pages

2.1: Proximate composition of pigeon pea (g/100g). 21

4.1: Microbial analysis of fermented and unfermented 38

sweet potato-pigeon pea weaning food

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 LIST OF FIGURES

Figures Pages

1: Preparation of Fermented sweet potato-pigeon pea flour 31

2: Preparation of unfermented sweet potato-pigeon pea flour 32

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 ABSTRACT

Microorganisms are one of major factors that affect the shelflife of any product especially
product with high moisture content and products that contain some nutrients which are needed
for their growth. The study was conducted to determine the microbial analysis of fermented and
unfermented sweet potato-pigeon pea weaning food (FSPWF and USPWF). Both products were
processed in the processing laboratory and the analysis was carried out in the chemistry
laboratory of Food Science and Technology. Microbial analysis contents of the formulated
blends were determined using standard methods. There were significant differences (p ≤ 0.05) in
the total viable count, coliform count but there was a significant difference between yeast and
mould count. The weaning foods prepared from fermented and unfermented pigeon pea and
sweet potato significantly improved the microbial properties of the food. The total viable and
mould counts were within the same range for both the fermented and unfermented weaning food
and are safe for human consumption when compared with <10^6cfu/g as recommend by
microbiological standards.

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 CHAPTER ONE

1.0 INTRODUCTION

Ten million children under the age of 5 years old die each year (Bryce et al., 2005). More

than half of the deaths occur because of malnutrition. If adequate health systems were in place

nearly 2/3 of the deaths could be prevented. Part of the health systems picture is to promote

appropriate feeding practices for infants and young children. If at all feasible breastfeeding is

recommended during the first six months, the most vulnerable period for developing under-

nutrition remains the transition from breastfeeding to family foods. Breast milk composition may

vary dramatically between women and from the beginning and month six (Allen et al., 2001).

Breast milk contains everything a newborn child needs to grow. Breastfeeding should continue

throughout gradual weaning process through at least the first year of life and for two years if

possible in order to provide the best nutrition for the growing child, with the exception of cases

where there may be the risk for HIV transmission from mother to child (Laskey et al., 2000).

Weaning is the process of transition from a breast feeding to a semi solid diet for the infant.

A reduction in breast milk consumption and the protection it provides during gastrointestinal

infection can increase the risk for illness due to diarrhea in children during weaning (Mata et al.,

2004). Poor quality of weaning foods and improper weaning practices predispose infants to

malnutrition, growth retardation, infection, diseases and high mortality (Onofiok et al., 2005).

The World Health Organization recommends a gradual weaning period from 6 months to 2 years

(WHO, 2006). This allows for the child to still receive the benefits from breastfeeding, while

also consuming the necessary nutrients from the complementary foods. Foods should be

prepared adequately containing the required nutrients as well as appropriately with a suitable

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texture and temperature (WHO, 2004). Without the knowledge of proper weaning practices as

well as a perception of the child’s hunger needs, malnutrition and illness may ensue. The

weaning period is therefore a vulnerable time when the child should be attentively cared for and

observed so as to maintain health (Dewery, 2001).

Sweet-potato (Ipomoea batatas), a sweet-tasting tuberous root, is ranked fourth in terms of

consumption in the world after wheat, maize, and rice (Odebode et al., 2008). Nigeria was

ranked the leading producer of sweet-potato in Africa between 1993 and 2013 (Olatunde et al.,

2016). It is rich in carbohydrate consisting mainly of starch and sugar (occurring as sucrose,

glucose, and fructose), and small amounts of pectins, hemicellulose, and cellulose (Preedy et al.,

2011; Saeed et al., 2012; Onabanjo and Ighere, 2014). Other chemical constituents of sweet-

potato include protein, dietary fiber, β-carotene, vitamins B, C, and E, and minerals such as

manganese, potassium, and iron. It is a beneficial food for the diabetics, as preliminary studies

on animals have revealed its ability to assist to stabilize blood sugar level and lower insulin

resistance (Odebode et al., 2008; Preedy et al., 2011).

Pigeon pea (Cajanus cajan) is an important underutilized legume in south-west Nigeria

(Fasoyiro and Arowora, 2013), where it is locally known as otili. It is tolerant to drought and has

wide adaptability to different environmental conditions (Troedson et al., 2000). It contains 20%–

22% of all essential amino acids particularly lysine and 18%–35% protein, and therefore

desirable in overcoming the incidence of protein-energy malnutrition in Nigeria (Okpala and

Okoli, 2011; Tiwari et al., 2011; Anuonye et al., 2012). Pigeon pea is rich in dietary minerals

such as calcium, copper, phosphorus, magnesium, iron, sulfur, and potassium, and water- soluble

vitamins such as thiamine, ascorbic acid, riboflavin, and niacin (Foodnet, 2002; Kaushal et al.,

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2012). It is a good source of slow release carbohydrates, making it a suitable raw material for the

formulation of low glycemic index food product (Morales-Medina et al., 2012).

The development of weaning foods based on locally available cereals and legumes has

been suggested by the Integrated Child Development Scheme (ICDS) and Food and Agriculture

Organization (FAO) to combat malnutrition among children of low socio-economic groups

(Imtiaz et al., 2011). During formulation of any weaning foods made from locally available raw

materials; the techniques of food preparation process, handling, storage, sanitation, sensory

properties, and food quality and safety issues in general should be taken in to account (Amuna et

al., 2000; Ifediora, et al. 2006). This is because; weaning foods prepared under unhygienic

conditions are shown to be heavily contaminated with pathogenic microorganisms (Nkere et al.,

2011). Contamination of food including drinking water with microbial agents is the major risk

factor in the transmission of diarrhoeal diseases in infants and young children. Contaminated

hands and cooking utensils contribute greatly to the contamination of weaning foods, especially

among mothers who do not observe proper hygienic conditions (Michaelsen et al., 2000).

1.1 Aim and Objectives of the Study

The aim of this project study is to examine the microbial analysis of fermented and

unfermented sweet potato-pigeon pea weaning food.

1.1.1 Objectives of the study

The objectives of the study include the following:

i. to produce fermented and unfermented sweet potato-pigeon pea weaning food

ii. to evaluate the microbial analysis of the samples

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 CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 Sweet Potato (Ipomoea batatas L)

Sweetpotato, Ipomoea batatas L. (Lam.), is an important economic crop in many

countries. In terms of annual production, sweetpotato ranks as the fifth most important food crop

in the tropics and the seventh in the world food production after wheat, rice, maize, potato,

barley, and cassava (FAO, 2016). Sweetpotato fulfills a number of basic roles in the global food

system, all of which have fundamental implications for meeting food requirements, reducing

poverty, and increasing food security (El‐Sheikha and Ray, 2017). Sweetpotato roots have high

nutritional value and sensory versatility in terms of taste, texture, and flesh color (white, cream,

yellow, orange, purple). The varieties with high dry matter (>25%), white‐cream flesh color, and

mealy firm texture after cooking are preferred by the consumers in the tropics. These varieties

are known as tropical sweetpotato (e.g., “bianito,” “batiste,” or “camote”). The purple‐fleshed

sweetpotato varieties with attractive color and high anthocyanin content are the specialty type in

Asia.

In the United States, the commercially popular type is the orange‐fleshed sweetpotato

with low dry matter content (18–25%), high β‐carotene level, sweet and moist‐texture after

cooking. This sweetpotato type is imprecisely called “yam,” which is not the true tropical yam of

Dioscorea species. Historically, African Americans in Louisiana referred this moist‐ sweetpotato

as “nyami” because it reminded them of the starchy tuber of that name in Africa. The Senagalese

word “nyami” was eventually shortened to the trademark “yam” popular in the United States.

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Commercial packages with “yam” labels are required by the US Department of Agriculture to

have the word “sweetpotato” in the label to avoid confusion to the consumers (Estes, 2009).

Depending on the flesh color, sweetpotatoes contain high levels of β‐carotene,

anthocyanins, phenolics, dietary fiber, vitamins, minerals, and other bioactive compounds. The

β‐carotene in orange‐fleshed sweetpotatoes can play a significant role as a viable long‐term food‐

based strategy for combating vitamin A deficiency in the world. Studies in Africa demonstrated

that increased consumption of orange‐fleshed sweetpotatoes improved the vitamin A status of

children, pregnant women, and lactating mothers (Van Jaarsveld et al., 2005; Low et al., 2007;

Hotz et al., 2012). Further, polyphenolics from purple‐fleshed sweetpotatoes exhibited diverse

biological activities, including protective antioxidant, anti‐inflammatory, anticancer, antidiabetic

and hepatoprotective activity effects (Lim et al., 2013; Hu et al., 2016). Sweetpotato has a strong

potential to contribute to better nutritional quality of our diets around the world.

2.1.1 Production and Consumption

Sweetpotato has wide production geography, from 40° north to 32° south latitude of the

globe, and it is cultivated in 114 countries. The world total production of sweetpotatoes was

106.60 million metric tons (MMT) in 2014. Since the mid‐2000s, global production has ranged

from a low of 101.28 MMT in 2007 to a high of 147.17 MMT in 1999. In 2014, about three‐

fourth of the global production was from Asia and Pacific Islands, followed by Africa with about

21%, while the Americas (North, Central, and South) account for about 3.6%. China was the

leading producer of sweetpotatoes, with 71.54 MMT or about 67% of the global production,

followed by Nigeria (3.78 MMT), Tanzania (3.5 MMT), Ethiopia (2.7 MMT), and Mozambique

(2.4 MMT). The United States was the tenth largest producer, with 1.34 MMT production. Only

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two countries in Europe, Portugal and Spain, grow sweetpotatoes, with 22,591 and 13,550 metric

tons produced in 2014.

In comparison to other major staple food crops, sweetpotatoes have good adaptability to

marginal growing conditions, short production cycle, and high yield potential. The average world

yield of sweetpotatoes is about 14 tons per hectare. Under subsistence conditions in many areas

of the tropics, the average sweetpotato yield is about 6 metric tons/hectare, far below the 20–26

metric tons/hectare obtained in China, Japan, and the United States, where improved varieties,

fertilizer applications, and cultural managements have been introduced. The per capita

consumption is highest in places where sweetpotatoes are consumed as a staple food, e.g., Papua

New Guinea at 550 kg per person per year, the Solomon Islands at 160 kg, Burundi and Rwanda

at 130 kg, and Uganda at 85 kg. The average annual per capita consumption of sweetpotatoes is

estimated at 18 kg in Asia, 9 kg in Africa, 5 kg in Latin America. Between 2000 and 2014,

sweetpotato consumption in the United States increased nearly 80%, from 1.9 kg to 3.4 kg per

capita (FAO 2015; Johnson et al., 2015).

Sweetpotato consumption has been greatly enhanced by the wide spread commercial

availability of frozen “French‐fried” sweetpotatoes. To accommodate this recent growth trend,

increased modern processing capacity has been built within the southern US sweetpotato

growing regions.

2.1.2 Classification and Origin

The sweet potato (I. batatas L.) is a dicotyledonous plant belonging to the morning glory

or Convolvulaceae family. It is a new world crop, though there is still some confusion that exists

regarding its origin, and primary and secondary centers of diversity. Roullier et al. (2013a,b) and

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Grüneberg et al. (2015) have published thorough reviews of this topic. In brief, using data from

morphology, ecology, and cytology, Austin (2008) has postulated that cultivated sweetpotatoes

originated somewhere in the region between the Yucatan Peninsula of Mexico and the mouth of

the Orinoco river in northeastern Venezuela. Recent studies conducted by Roullier, et al. (2013

a, b) incorporating chloroplast DNA and molecular phylogeny analyses confirm this hypothesis.

They also suggest that I. batatas most likely evolved from at least two distinct

autopolyploidization events in wild populations of a single progenitor species most likely I.

trifida.

Secondary contact between sweetpotatoes domesticated in Central America and in South

America, from differentiated wild I. batatas or I. trifida populations, could have led to further

introgression. Molecular marker analyses conducted by Huang and Sun (2000) and Zhang et al.

(2000) also places Central America as the region with the most genetic diversity and probable

origin (Huang and Sun, 2000; Zhang et al., 2000). Remains of dried sweetpotato roots found in

Peru have been radiocarbon dated back to 8,000–10,000 years old, though it is unknown if these

were collected from the wild or were domesticated (Engel, 2000). Regardless of the center of

origin, sweetpotato was widely established in tropical regions of the new world around 2500 b.c.

(Austin, 2008). It was established in Polynesia, prior to European arrival (Roullier et al., 2013b).

Europeans in the 1500s spread the sweetpotato to Africa and India, with it arriving in China prior

to 1600. Secondary centers of diversity include New Guinea, the Philippines, and parts of Africa

(Roullier et al., 2013a, b).

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2.1.3 Postharvest Handling Practices

Storage

In temperate regions where production is limited to a summer season and marketing is

continuous, sweetpotatoes are stored year round. Varieties have been selected for both low

respiration and low water loss, giving a storage life up to 13 months or until the next crop is

harvested. Careful handling of sweetpotatoes is critical to ensure long‐term storage. Bruising and

skinning in the field are minimized by hand harvest or by using a combination of mechanical and

hand harvesting. Roots exposed to bright sun for more than 30 minutes may have a darkening of

skin called sun scalding, which is a cosmetic defect but can also be a site for postharvest decay.

Roots should not be harvested when the weather is too cold. Chilling injury is a function of

temperature and duration of exposure. Temperatures below 10°C will cause chilling, though

cooler temperatures will cause more damage. Chilling injury may not be seen for weeks after the

chilling occurs and can be expressed by various symptoms including increased respiratory rate,

greater susceptibility to decay, surface pitting, internal breakdown, hardcore and reduced

culinary quality (Truong et al., 2018).

After harvest, roots are immediately “cured” at 29–33°C and 85–90% RH with proper

ventilation for 4–7 days. Curing heals wounds that occur during the harvest, first by a

lignifications beneath cells damaged at harvest, and second by the formation of a wound

periderm beneath the lignified cells in a process called suberization. The healing provides a

pathogen barrier and reduces desiccation at the wound site resulting in less weight loss during

storage. Uncured roots do not store well but properly cured roots stored at 13–15°C and 85–95%

relative humidity will be marketable for up to 12 months (Edmunds et al., 2008). Good airflow is

17
essential to maintain oxygen and carbon dioxide exchange and allow for heat transfer. Cultivars

vary tremendously as to how long they will store and maintain the necessary quality. Curing also

produces changes in the culinary characteristics increasing moistness and sweetness (Walter,

2007).

Sweetpotatoes continue to respire during storage, converting starch to sugar, which is

then oxidized to carbon dioxide and water providing energy for the living cells. Over time, the

loss of dry matter will cause pithiness, a textural defect caused by an increase in intercellular

space, up to the point where there are air pockets in the root tissue. This is greatly accelerated by

warmer temperatures. Once temperatures go above 16°C, the roots will begin to sprout which

greatly increases the respiration rate and weight loss (Edmunds et al. 2008). Large commercial

storage facilities in developed nations can maintain very precise conditions to optimize root

storability and quality. In developing countries, storage of sweetpotatoes has been done for

hundreds of years and is still practiced using various pit, or underground storage structures. The

success of these structures depends on how close they come to maintaining the ideal temperature,

moisture, and oxygen levels as described. Storage losses due to rodents, weevils, and rots tend to

be high, and the length of time often limited to a few months (Truong et al., 2018).

Packing and Shipping

Market requirements, especially shape and size requirements, for sweet potato vary by

region. Where it is a subsistence food, shape, and size are not as important, but where it is a

luxury item, appearance is very important. In the United States, highly mechanized packing lines

are used to grade for strict size and shape parameters. Lines typically start with a tank of water

into which roots are dumped. This wets the roots for washing and allows roots to be metered

18
onto a conveyor system. Roots go through water rinse to remove soil, followed by an eliminator

to remove trash and small, unmarketable roots, usually accomplished by going across a set of

rollers at a specified width. Roots are then sorted, usually by hand, to remove decaying or

otherwise unmarketable roots. Roots that will be shipped for retail are then generally treated with

a fungicide to reduce decay. This is followed by sizing into various classes, some by diameter, or

with electronic sizers measuring both length and diameter. Roots are put into boxes, and boxes

onto pallets for efficient handling (Truong et al., 2018).

Bruising on packing lines can greatly affect shelf‐life of the sweetpotatoes, and care

should be taken in design and setup of the packing lines to reduce any impacts. The dump tank

drop‐ offs and onto conveyors, turns, and packing‐line speed and length account for much of the

damage and should be minimized (Edmunds et al., 2008). Market life, which begins when roots

are removed from bulk storage bins, of a sweetpotato is generally 2–3 weeks. The most common

disease in storage and packed sweetpotatoes is Rhizopus soft rot caused by the fungus Rhizopus

stolonifer. Present in most stored sweetpotatoes, it will contaminate packing lines and enter

through wounds produced during packing. Sanitation and minimizing wounds on packing lines is

the most effective control, and the main reason for the fungicide treatment. Care must be taken to

ensure that shipping containers are maintained at 13°C to prevent excessive respiration or

chilling damage (Truong et al., 2018).

2.1.4 Nutritional Composition of Sweet potato

All the plant parts, roots, vines, and young leaves of sweetpotatoes are used as foods,

animal feeds and traditional medicine around the world (Mohanraj and Sivasankar, 2014). In

Asia and Africa, the sweetpotato leaves are eaten as green vegetables. The nutrient content of

19
sweetpotato leaves varies among the varieties, harvest dates, crop years and cooking methods.

On dry weight basis, sweetpotato leaves contain 25–37% protein, 42–61% carbohydrate, 2–5%

crude fat, 23–38% total dietary fiber, 60–200 mg/100 g ascorbic acid, and 60–120 mg/100 g

carotene (Sun et al. 2014). They are also rich in calcium (230–1,958 mg/100 g), iron (2–22 mg),

potassium (479–5,230 mg), and magnesium (220–910 mg). The high level of phenolics (1.4–

17.1 mg/100 g dry weight), anthocyanins, and radical‐scavenging activities in sweetpotato leaves

indicates their potential benefits on human health and nutrition (Islam, 2006, Truong et al.,

2007).

Sweetpotato greens are very rich in lutein, 38–51 mg/100 g in fresh leaves, which are

even higher than the lutein levels in the vegetables that are known as a source for lutein, such as

kale (38 mg/100 g) and spinach (12 mg/100 g) (Menelaou et al., 2006). Novel galactolipids were

recently isolated and characterized from sweetpotato leaves (Napolitano et al., 2007), indicating

that this leafy vegetable can be a potential source of omega‐3 polyunsaturated fatty acid. Health

benefits and disease prevention of bioactive compounds in sweetpotato leaves have been

reported (Johnson and Pace, 2010). The nutrient composition of sweetpotato roots varies widely,

depending on the cultivar, growing conditions, maturity, and storage. Overall, sweetpotato roots

have a high moisture level with an average dry matter content of 25–30%. A wide range of dry

matter content of 13–45% from a sweetpotato germplasm collection was reported by Tsou and

Hong (2002) and Brabet et al. (2008).

Sweetpotato roots are good source of carbohydrates and generally low in protein and fat.

Protein content ranged from 1.73–9.14% on dry weight with substantial levels of nonprotein

nitrogen (Yeoh and Truong, 2006). Sweetpotato protein overall, however, is of good quality, and

the levels of essential amino acids compare significantly to the FAO reference protein (Maloney

20
et al., 2014). Most of the dry matter in sweetpotatoes consists of carbohydrates, primarily starch

and sugars and to a lesser extent pectins, cellulose, and hemicellulose. Dietary fiber in

sweetpotato roots range from 2 to 4% of fresh weight. Residues from sweetpotato starch and

juice processing of commercial varieties are good sources of dietary fiber, 16–36% of dry weight

(Mei et al., 2010; Truong et al., 2012a). Starch comprises 60–70% of the total dry matter, but the

values vary for different types of cultivars. As with other starches, sweetpotato starch granules

are made up of amylose (20%) and amylopectin and pasting temperatures are usually in a range

of 60–76°C (Zhu and Wang, 2014).

A special sweetpotato cultivar in Japan named Quick Sweet has starch gelatinization

temperature of <50°C and short cooking time. Short amylopectin chain length and cracking on

the hilum of starch granules contribute to the lower pasting temperature of the Quick Sweet

cultivar (Takahata et al., 2010). Much variability in sugars exists between sweetpotato types.

Truong et al., (2006) found total sugars to vary from 5.6% in a Filipino cultivar to 38% in a

Louisiana cultivar on a dry weight basis (db). Sucrose, glucose, and fructose make up the

majority of the total sugars in raw sweetpotato roots. During cooking, amylases act on the

gelatinized starch resulting in the formation of maltose in cooked sweetpotatoes. There is

substantial genetic diversity within the sweetpotato genotypes collected around the world in term

of sugar content and degree of sweetness that contribute to the consumer preferences of

processed products (Kays et al., 2005; Leksrisompong et al., 2012). The glycemic indices of

cooked sweetpotatoes were about 63–66, indicative of moderate glycemic index food (Allen et

al., 2012).

Ash content of sweetpotatoes is approximately 3% of the dry weight or between 0.3%

and 1.0% of the fresh weight basis (fwb). Potassium is the mineral with the greatest

21
concentration in sweetpotato, with an average of 396 mg/100 g fwb. Phosphorous, calcium,

magnesium, iron, copper, and magnesium are also present in significant amounts (Woolfe 2002).

Sweetpotato roots also contain vitamins such as ascorbic acid, thiamin (B1), riboflavin (B2),

niacin (B6), pantothenic acid (B5), folic acid, and vitamin E. Bradbury and Singh (2006)

reported values between 9.5 and 25.0 mg/100 g (fwb) for ascorbic acid and 7.3–13.6 mg/100 g

(fwb) for dehydroascorbic acid resulting in a total vitamin C range of 17.3–34.5 mg/100 g for the

sweetpotato roots. Orange‐fleshed sweetpotatoes are rich in β‐carotene. A wider range of β‐

carotene content in cooked orange‐fleshed sweetpotatoes, 6.7–16.0 mg/100 g fwb, has been

reported by different investigators (Bovell‐ Benjamin 2007).

The sweetpotato carotenoids exist in an all trans configuration, which exhibits the highest

provitamin A activity among the carotenoids. Van Jaarsveld et al. (2005) advocate the increased

consumption of orange‐fleshed sweetpotatoes as an effective approach to improve the vitamin A

nutrition in the developing countries. Total carotenoid content is correlated with the dry matter

content and sensory attributes involving visual, odor, taste and textural characteristics of cooked

sweetpotatoes. Doubling in carotenoid content would result in a decrease of about 1.2% of dry

matter content in sweetpotato varieties (Tomlins et al., 2012). Epidemiological studies indicated

the beneficial effects of high carotene diets in reducing the risks of cancer, age‐related macular

degeneration, and heart diseases (Tanumihardjo, 2008).

2.1.5 Processing and Utilization

Sweetpotato roots and other plant parts are used as human food, animal feed, and

processing industry. For industrial processing, starch, sugars, and natural colorants are the major

intermediate products that can be used in both food and nonfood processing industry.

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Sweetpotato varieties with high levels of dry matter (35–41%), total starch (25–27%), and

extractable starch (20–23%) are available for starch processing (Brabet et al., 2008). There are

many small and medium factories in Asia producing about 26% of starch production (Bovell‐

Benjamin, 2007). The process for manufacturing sweetpotato starch is basically similar to the

starch extraction from other sources. The roots are ground in limewater (pH 8.6–9.2) to prevent

browning due to polyphenol oxidase, to dissolve pigments, and to flocculate the impurities. The

extracted starch is separated from the pulp by thoroughly washing over a series of screens,

bleaching with sodium hypochlorite, and then settling by gravity or centrifugation. In small‐

scale establishments, starch is stored wet in concrete tanks or sun‐dried to a moisture content of

about 12%, pulverized and screened. Centrifugation and mechanical drying, such as flash dryer,

are commonly used for medium‐ scale factories.

Sweetpotato starch is used in the production of traditional noodles, vermicelli, thickening

agents, or converted into sugar syrups, which are used in many processed food products. The

sweetpotato starch and sugars are also utilized in the production of fuel alcohol, monosodium

glutamate, microbial enzymes, citric acid, lactic acid, and other chemicals (Padmaja, 2009). In

Japan, the orange‐ and purple‐fleshed sweetpotatoes have been used in commercial production of

natural beta‐carotene and anthocyanin pigments in beverages and other food products. The

following sections describe recent developments in processing of sweetpotatoes into functional

ingredients and common food products.

23
2.1.6 Health Benefits of Sweet Potato

 Antioxidative and Antimutagenicity

Cancers occur through such processes as initiation, promotion and progression in body

cells. Initiation is a kind of mutation that occurs in cancer and anticancer genes. Thus, controlling

the gene mutation brought about by the carcinogens leads to cancer prevention. Sweetpotato

leaves are a good supplementary resource of antioxidants and antimutagenic compounds. An

investigation was conducted to examine the effects of 82 kinds of vegetable juice and plant

components on the division and multiplication of cancer cells, and it was found that sweetpotato

has especially high cancer checking rates (Haliru et al., 2015).

 Anti-diabetes

The latter stage of non-insulin dependent diabetes mellitus (NIDDM), one of the major

diseases of adults, is caused by the decrease in the secretion of insulin by the pancreatic

Langerhans cells. Prevention of NIDDM and inhibition of its serious side effects such as

retinopathy, neuropathy and cataracts are important subjects for researchers. The current diabetes

mellitus population in the United States is increasing and is estimated at more than 18 million

persons. Diabetes contributes to the death of more than 213,000 Americans each year and is also

a leading cause of heart disease, blindness and kidney failure. Foods with antidiabetic effect are

desired for diet therapy. Several researchers report that sweetpotato leaves have antidiabetic

compounds that reduce blood glucose content significantly in model rats (Li and Kao, 2000).

24
  Antibacterial Activity

Lyophilized sweetpotato leaf powder from the variety ‘Simon1’ strongly suppressed the

growth of O157, and its effect was detectable even after autoclave treatment. The anti-bacterial

extract revealed that the main components were polysaccharides. Furthermore, the water

extracted from the leaves suppressed effectively the growth of other foodpoisoning bacteria such

as Staphylococcus aureus and Bacillus cereus as well as pathogenic E. coli.

2.2 Pigeon Pea (Cajanus cajan (L) Millsp)

Pigeon pea (Cajanus cajan (L) Millsp). (In Sanskrit: Adhaki, Hindi: Arhar, English:

Pigeon pea, Bengali: Tur) is a perennial member of the family Fabaceae. It is also known as red

gram, congo pea, gungo pea, and no-eye pea (Wu, 2009). Pigeon pea is a tropical and subtropical

species particularly suited for rainfed agriculture in semi-arid areas due to its deep taproot, heat

tolerance and fast growing habit (Mallikarjuna et al., 2011). It is the sixth most important grain

legume crop grown in the semi-arid tropics of Asia, Africa and the Caribbean under a wide

variety of cropping systems (Mula and Saxena, 2010). Pigeon pea is a legume reported to contain

20-22% protein, 1.2 % fat, 65% carbohydrate and 3.8% ash (FAO 2002). Its demand in India is

significant because it can provide high quality protein in diet, especially to the vegetarian

population (Bhattacharjee et al., 2013). It is a fast growing, hardy, widely adaptable and drought

resistant (Bekele-Tessema, 2007). It is very heattolerant and grows better in places where

temperatures range from 20° to 40°C and which are deprived of frost (FAO, 2016).

Being environmental friendly by fixing nitrogen, flexibility for mixed cropping or inter

crop, due to this it has significant position in dry land farming systems especially adopted by

small and marginal farmers in many parts of world (Pandit et al., 2015). In addition to being

25
efficient in fixing nitrogen in field conditions, pigeon pea rhizobia also present other

biotechnological applications, such as biopolymer production and enzymatic activity (Junior et

al., 2011; Fernandes et al., 2012). Its deep taproot is able to extract nutrients (like P) from the

low layers lower layers of soil, and bring them to upper layers where they can benefit to other

crops (Valenzuela, 2011). Thanks to drought resistance as it can be considered of utmost

importance for food security in regions where rain failures are prone to occur (Crop Trust, 2014).

2.2.1 Origin of Pigeon Pea

The centre of origin of pigeon pea is probably peninsular India, where the closest wild

relatives (Cajanus cajanifolia) occur in tropical deciduous woodlands (Van der Maeson., 2005).

But, some other opinions state that the origin of Cajanus cajan was either from Northeastern

Africa or India (Ecocrop, 2016). Archaeo logical finds of pigeon pea dating to about 3400 years

ago (14th century BC) have been found at Neolithic sites in Karnataka and its border areas

(Tuljapur-Garhi in Maharashtra and Gopalpur in Orissa) and also the south Indian states such as

Kerala, where it is called Tomara Payaru (Fuller and Harvey, 2006). From India it traveled to

East Africa and West Africa. There, it was first encountered by Europeans, so it obtained the

name Congo Pea. By means of the slave trade, it came to the American continent, probably in the

17th century (Carney and Rosomoff, 2009). According to Van der Maesen, 2000, India is the

native of pigeon pea because of its natural genetic variability available in the local germplasm

and the presence of its wild relatives in the country.

2.2.2 Botany

Pigeon pea plants are a perennial legume which can reach a height upto 3 feet to 12 feet.

The compound leaves of this plant consist of three green leaflets which have a pubescence and

26
darker color on the upper side and a graygreen color on the underside. The flowers are yellow

with red lines or completely red on the exterior (Jonael Bosques). The flowers are bisexual,

zygomorphic and predominantly yellow (Sundaraj and Thulasidas, 2000). The inflorescence is

raceme which contain up to ten flowers per panicle and usually two flowers open at a time on a

single inflorescence (Sharma and Green, 2000). The androecium has 10 stamens bunched into

two groups (diadelphous) of 9 and a single free stamen that is attached at the base of androecium.

The fruit of pigeon pea is a flat, straight, pubescent, 5-9 cm long x 12-13 mm wide pod. It

contains 2-9 seeds that are brown, red or black in colour, small and sometimes hard coated

(FAO, 2016; Bekele-Tessema, 2007).

The 100 seed weight of short duration cultivated varieties are low (generally 6-8 grams)

as compared to long duration varieties (9-13 g). Root of pigeon pea is well developed in upper 60

cm soil profile (Natarajan and Willey, 2000). The root system consists of fine lateral roots as

well as a large taproot. This root provides the capacity to overcome moderate drought periods

(Jonael Bosques). The vegetative growth of the pigeon pea starts slowly, but accelerates when

the plant reaches 2 to 3 months.

2.2.3 Nutritional composition of pigeon pea

Proximate composition: The proximate compositions such as moisture, protein, fat,

carbohydrate, ash and dietary fiber of pigeon pea is shown in Table 1 (Eltayeb et al., 2010;

Olalekan and Bosede, 2010; Adamu and Oyetunde, 2013; Kunyanga et al., 2013). The pigeon

pea seed is made up of 85% cotyledon, 14% seed coat, and about 1% embryo. It is a rich source

of protein, carbohydrates, minerals and vitamins. It’s protein content ranges between 20-22%

27
CHO between 51.4 – 58.8%, Crude fibre between 1.2 – 8.1% and lipid between 0.6 – 3.8% (Oke,

2014).

Carbohydrates: The starch and non-starch are the major constituents of carbohydrates of pigeon

pea consisting of significant amount of α-galactosides. Different carbohydrate fractions of

pigeon pea are (i) available carbohydrates which are digested in the small intestine and (ii)

unavailable carbohydrates like oligosaccharides, resistant starch, non cellulosic polysaccharides,

pectins, hemicelluloses, and cellulose, which are not digested in the small intestine (Cummings

and Stephen, 2007).

The unavailable carbohydrates of pigeon pea like fructans and raffinose family

oligosaccharides (RFOs) are the major water soluble carbohydrates which adversely affect

bioavailability of certain vital nutrients (Apata, 2008; Devindra et al., 2012).

Glycemic carbohydrate: Carbohydrates which provide glucose for metabolism is referred to as

‘glycaemic carbohydrate’, whereas carbohydrates that reaches the large intestine prior to being

metabolized, are called as ‘nonglycaemic carbohydrate’. Most of the available carbohydrates,

some oligosaccharides and rapidly digested starches may be classified as a glycaemic

carbohydrate (Cummings and Stephen, 2007). In a study cooked legumes including pigeon pea,

chickpea, black gram, mung bean and white bean were tested for blood glucose response among

healthy human subjects. The glycemic response to pigeon pea was 30.99 (Panlasigui, 2009).

Recently Devindra et al. (2016) have reported the lower glycemic index of pigeon pea among the

commonly consumed legumes tested.

Amino acid profile: Generally the sulphur containing amino acids (methionine and cystine) are

limiting in pulses. Pigeon pea seeds contain high amounts of lysine, leucine, aspartic acid,

28
glutamic acid and arginine and provide essential amino acids when consumed with cereals and

other sulphur containing amino acids (AdeOmowaye et al., 2015). In a study Pigeon pea can

replace soybean without adversely affecting the performance of rabbits (Adamu and Oyetunde,

2013).

29
 Table 2.1: Proximate composition of pigeon pea (g/100g).

Oke, Eltayeb et al. Eltayeb et al. Adamu, Kunyunga et al.

Proximate
(2014) (2010) (2010) (2013) (2013)
Moisture 11.20 8.0 8.45±0.95 0.24 11.27±0.04
Protein 22.40 21.0 24.46±0.32 30.53 17.95±0.06
Fat 2.74 1.7 4.78±0.22 3.68 2.77±0.57
Carbohydrat
48.19 63.6 56.63±0.48 50.08 57.45±0.04
e
Fibre 7.25 2.5 1.10±0.10 5.54 6.98±0.08
Ash 8.22 3.2 4.58±0.40 9.93 3.58±0.21

30
Fatty Acid Profile: The major saturated fatty acid in pigeon pea is the palmitic acid which

constitutes 15-25% in the neutral lipids, 20-40% in the glycolipids, and 26-30% in the

phospholipids. A study was conducted to know the nutritional potential of nine underexploited

legumes in Southwest Nigeria. The most abundant polyunsaturated fatty acid (PUFA) identified

in pigeon pea was linoleic acid (C18:2) (Ade-Omowaye et al., 2015). Caprylic, lauric, oleic and

eicosanoic acids were present only in small quantities. The resistant starch derived from

processed red gram showed a higher amount of short chain fatty acids. Whereas Indian food

composition tables reported the palmitic acid content of pigeon pea as 236 ± 11.0, oleic acid was

78.55± 6.71, stearic acid was 40.95± 3.31 mg/100g respectively (Longvah et al., 2017).

Vitamin and mineral content: Pigeon pea is a good source of water soluble vitamins, like

thiamine, riboflavin, niacin etc. Pigeon pea was reported to be the richest vegetable for Vitamin

C with 569 mg/100g. Its content was three times higher than for peppers and could cover more

than 949% of the recommended daily intake (RDI) of Vitamin C and pigeon pea was found to

have high carotenoid content with 364.3 µg/100g when compared to other pulses (Ellong et al.,

2015). Whereas Indian food composition tables reported the thiamin content of pigeon pea as

0.74 , riboflavin 0.15, niacin, panto thenic acid 1.56, total B6 0.42,biotin 0.65, total folates 229

mg/ 100g respectively (Longvah et al., 2017).There are limi ted studies on the lutein and

zeaxanthin levels in pigeon pea.

Pigeon pea is a good source of minerals such as phosphorus, magnesium, iron, calcium,

sulphur and potassium but low in sodium (Kunyanga et al. 2013). Sangle, (2015) have reported

minerals constituents of two viable mutant varieties of pigeon pea. Mean content of nitrogen

31
ranged from 1.95%- 3.33% and 2.24% - 3.17%, calcium content ranged from 0.25% - 0.37% and

0.26% - 0.51% and phosphorus content of viable mutants ranged from 0.56 % - 0.72 % and

0.58% - 0.80% in varieties of pigeon pea respectively (Sangle, 2015).Whereas Indian food

composition tables reported the calcium content of pigeon pea as 1.39, copper 1.32, iron 5.37

mg/100g respectively (Longvah et al. 2017).

2.2.4 Health benefits of Pigeon Pea

The nutritional components of pigeon pea are widely considered crucial for human

nutrition, because of phytochemicals, bioactive compounds which play vital roles in humans.

Traditionally Cajanus cajan leaves have been used by Rabha tribe to cure jaundice and also

described as useful for the treatment of smallpox, chicken pox, measles and also as an astringent,

mouthwash by local people of North East India (Sarma et al., 2015). Pigeon pea can be a source

of remedy in the control of sickle cell anaemia. In India, many Sickle cell disease (SCD) patients

are using pigeon pea for effective management of erythrocytes sickling in Chhattisgarh (Verma,

2015). In an ethnomedical survey pigeon pea plant was recorded as traditional medicinal plant

used in Northern and South-Eastern Côte d’Ivoire for the treatment of anaemia (Kone et al.,

2011). Pigeon pea also has anti-ulcer potential (Mansoor et al., 2015).

The European Society of Hypertension reco mm ends diet and lifestyle approaches as a

basis for prevention and treatment of hypertension. From animal model, it was both water

extracts of pigeon pea and water extracts of B. subtilis fermented pigeon pea improved systolic

blood pressure and diastolic blood pressure in spontaneously hypertensive rats (Lee et al., 2015).

Prebiotic potential: A prebiotic is defined as a selectively fermented ingredient that allows

specific changes, both in the composition and/or activity in the gastrointestinal microflora that

32
confers benefits upon host well-being and health (Roberfroid et al. 2010). Fermentation of

dietary carbohydrates provides the energy for the growth and activity of the intestinal microbiota.

Several studies established the role of prebiotics in weight control, by improving microbial

balance, ameliorating adiposity and increasing mucosal integrity with decreased inflammation

(John et al., 2012). A number of mechanisms have been implicated in the link between intestinal

microbiota, increased fatty acid metabolism, and storage of calories as fat (Mallappa et al.,

2012). The prebiotics can be a tool to modulate gut microbiota which plays a role in the

pathophysiology of obesity.

A diet rich in non-digestible carbohydrates induced significant weight loss and

concomitant structural changes of the gut microbiota in simple obese children (Zhang et al.,

2015). In another study consumption of prebiotics for 16 weeks significantly improved

bifidobacterial abundance in overweight and obese children and proved that prebiotic fiber is a

potential treatment option to reduce body fat by gut microbiota modulation (Nicolucci et al.,

2015). Recently Devindra et al. (2017) have demonstrated the prebiotic potential of red gram

raffinose oligosaccharides in an animal model. The results of the prebiotic potential of red gram

oligosaccharides have shown a hypolipidemic effect and lowered blood glucose level, improved

HDL and decreased LDL. This new array of research may provide enough evidence for the

prebiotic potential of pigeon pea.

Antioxidant potential: The bioactive compounds of pigeon pea seeds contain some defense

machinery. Four important compounds, pinostrobin, cajaninstilbene acid (CSA), vitexin and

orientin isolated from ethanolic extracts of pigeon pea found to possess significant antioxidant

activities (Pal et al., 2011). Antioxidant potential of pigeon pea seed husk was investigated and

results revealed a potent anti-oxidant activity (Rani et al., 2014). The aqueous extract of the

33
pigeon pea had the highest antioxidant activity possibly because of the presence of polyphenols

(Mahitha et al., 2015). Uchegbu and Ishiwu (2015) studied antioxidant activity of extract of

germinated pigeon pea in alloxan-induced diabetic rats. Consumption of germinated pigeon pea

extract resulted in reduction of fasting blood glucose level and LPO in diabetic rats (Uchegbu

and Ishiwu, 2015).

Hypocholesterolemic effect: The consumption of pulses and legumes has been reported to

ameliorate serum cholesterol levels and increase the saturation levels of cholesterol in the bile.

Pigeon pea is a good source of saponins which have been implicated for the control of high

cholesterol level and they bind to the bile salts (Aja et al., 2015).

Overweight and obesity: The diet high in whole grains is associated with the lower body mass

index, smaller waist circumference, reduced risk of overweight and obesity (Butt et al., 2011).

Pigeon pea has high protein content and this dietary protein may promote weight loss by

inducing satiety hence decrease energy expenditure. Using data from the National Health and

Nutrition Examination Survey (NHANES), it was observed that adults who consumed a variety

of legumes had significantly lower body weights compared with those who did not consume

legumes (Polak, 2015).

Role in diabetes (Hypoglycemia): It is acknowledged that certain fibre rich foods like whole

pulses can affect glycemic control in diabetes and hyperlipidemia. Pigeon pea is the most

effective hypoglycemic medicinal plant commonly studied in relation to diabetes and their

complications and owing to its varied degree of hypoglycemic activity. In a study substitution of

red meat with legumes improved lipid profiles and glycemic control among diabetes patients

(Hosseinpour-Niazi, 2015).

34
 In another study, there was a reduction in glucose levels of rats treated with ethanol

extract of pigeon pea leaves and an increase in ALT, AST and ALP levels when compared with

those treated with Moringaoleifera extract (Aja et al., 2015b). In a study, crude methanol extract

of pigeon pea seed husks mitigated starch-induced postprandial glycemic excursions and reduced

glycemic load in rats similar to the standard drug acarbose (Tiwari et al., 2013). The hypogl

ycemic effect of crackers produced from sprouted pigeon pea caused hypoglycemic effect in

diabetic rats and led to a reduction of measured biochemical indices (Uchegbu, 2016).

Cancer prevention: Alkaloids of pigeon pea have been implicated for inducing a stress response

and apoptosis in human breast cancer cell. DNA fragmentation was observed in human breast

cancer cells treated with cajanol, a novel anticancer agent from pigeon pea roots (Luo et al.,

2010). The methanol extract of the plant has been shown to demonstrate cytotoxicity against

three cancer cell lines, namely human breast adenocarcinoma cell line MCF-7, human large cell

lung carcinoma cell line COR-L23 and human amelanotic melanoma C32 (Ashidi et al., 2010).

Antimicrobial effect: Pigeon pea plant extract is inhibitory to some bacterial pathogens (Braga

et al., 2007). The leaf part of pigeon pea is an excellent source of natural antimicrobial

substances. Moreover, cajanuslactone is a potential anti-bacterial agent against Gram-positive

microorganisms (Kong et al., 2010). The presence of tannins, flavonoids and alkaloids in pigeon

pea extract has clinically relevant antifungal activity (Brito et al., 2012).

A study investigated the protective effect of cajaninstilbene acid against corticosterone

induced injury in PC12 cells and examined the potential mechanisms for the same (Jiang et al.,

2014). Different extracts of leaf, seed and root of the pigeon pea were proven to be a great

potential source of antibacterial compounds (Devi et al., 2016) and also aid in wound healing

35
activity. In a study, wound healing activity of hydrogel obtained from pigeon pea seed husk was

carried out in albino rats. Gel formulation showed significant antibacterial activity against both

gram positive and gram negative selected bacteria and the percentage wound closure and

epithelialization for the gel formulation treated group was comparable with those of standard

group treated with Band aid (Patil and Mastiholimath, 2011).

Hepatoprotective: Pigeon pea is already known for its hepatoprotective function. Pigeon pea

plant protein extracts can up-regulate and counteract the inflammatory process, minimize the

damage to the liver, delay disease progression, and reduce its complications in liver (Rizk et al.,

2014) and kidneys (Aly et al., 2016). The pigeon pea extract was potential towards antioxidative

protection against ironoverload-induced liver damage in mice and improved hepatic antioxidants

(Sarkar et al., 2013).

Pigeon pea plant extract also had promising anthelmintic effects against F. hepatica

(Alvarez et al., 2015) and a poly herbal formulation from pigeon pea could protect the liver cells

from CCl4 - induced liver damages (Arka et al., 2015). In another study hepatoprotective activity

of pigeon pea was studied in sodium fluoride treated Swiss albino (BALB/C) mice. Treatment

with extract of pigeon pea exhibited significant anti-oxidant and hepatoprotective activity

(Kayathri et al., 2015).

2.2.5 Use as functional foods

A study was conducted to investigate the survival of Lactobacillus reuteri ATCC 55730

in creams, prepared with pigeon peas and oat. The study concluded that L. reuteri ATCC 55730

had the highest viability in cream with 40% pigeon pea and 20% oat (Barboza et al., 2012). The

effect of the use of pigeon pea as a substrate in the production of a legume-based fermented

36
product with Lactobacillus acidophilus ATCC 314 or Lactobacillus casei ATCC 393 was

studied by Parra et al. Pigeon pea-based fermented probiotic product was suitable for both the

strains (Parra et al., 2013). In a study, the effects of a dietary prebiotic, inulin and probiotic was

investigated in mice using cellulose-based AIN-93G diets under conditions allowed for the

growth of commensal bacteria (Kuo et al., 2013).

2.2.6 Socio-economic Impact of Pigeon Pea

Pigeonpea is widely recognized for its agronomic and nutritional benefits to rural

households, including its capacity to earn foreign exchange for many African countries, such as

Kenya, Tanzania, Malawi and Uganda (Shiferaw et al., 2005). Studies carried out to date are

now focused on improvement of productivity and quality of production, but little is said about its

role in improving the nutrition and livelihoods of farmers (ICRISAT, 2001; Jones et al., 2002;

Mergeai et al., 2005). Moreover, this limited attention has focused on the preferences of farmers

for consumption and the motivational factors causing this.

A recent study highlights how pigeonpea could promote rural development in Africa.

However, this study was based on a literature review and some statistical data from FAO. The

data from FAO regarding pigeonpea are not reliable as data for Mozambique are not included.

Thus, there is a need to carry out empirical research to study the real impact of pigeonpea on

small farmers. Furthermore, a project was implemented and reported in Uganda to address

handling, processing and marketing of pigeonpea in this country, but once more this was focused

on the marketing of the crop (Agona and Muyinza, 2004). Another study done in Kenya focused

on the incorporation of pigeonpea into the farming system, varieties used and its marketing, the

37
major constraints to production and provides some recommendations for investigations to

improve pigeonpea production (Mergeai et al., 2005).

Even though Shiferaw et al. (2007) did follow a socio-economic approach to study the

impact of improved pigeonpea varieties in Tanzania, it is again related to particular varieties that

were introduced by ICRISAT to alleviate the agronomic problems posed by pigeonpea

production and foster cash provision in the country. The available reports are more focused at

macro-level benefits. Hence, there has been no research executed at the micro-level, which draws

attention to how farmers, the producers of pigeonpea, are using it and its impact on their

livelihoods.

The approach employed in the current research to assess the socio-economic impact of

pigeonpea in Mozambique is the Sustainable Livelihoods Approach (SLA) which includes all

aspects of livelihood resources of small farmers and social relations. This study has used the

assets outlined within the SLA that include natural, human, physical, financial and social. These

are important as they are the foundations of any strategy that is adopted by the household to

ensure its members’ welfare. Additionally, the SLA focus is on the producers of pigeonpea, as it

was primarily developed with a focus on poor people. Moreover the approach is for the

promotion of rural development and poverty reduction. The impact of pigeonpea’s promotion on

livelihoods assets is analyzed, as a way of agriculture contributing to the broad process of

poverty alleviation. Additionally, by improving access to the capitals, farmer households can

progress along the scale of rural development and further steps to reduce poverty are more

certain.

38
 CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Materials

Sweet potato (Ipomoea batatas) and Pigeon pea (Cajanus cajam) used in the research

work was purchased from a local market in Owo, Ondo State. The samples were processed in

Food Processing Laboratory, Rufus Giwa Polytechnic Owo, Ondo State, Nigeria

3.2 Methods

3.2.1 Preparation of fermented sweet potato-pigeon pea flour

Fresh sweet potato root were washed, peeled, sliced and pigeon pea seeds were

winnowed, sorted, drained. The two cleaned samples (sliced sweet potato root and washed

pigeon pea were soaked together i.e fermented together for 72 hours. After 72 hours the

fermented samples it was washed, wet milled, sieved, sedimented for 24 hours, drained, sun

dried for 3 days and dry milled into powder, sieved and packaged in airtight container for further

analysis (Fig. 1).

3.2.2 Preparation of unfermented sweet potato-pigeon pea flour

Fresh sweet potato root were washed, peeled, sliced and pigeon pea seeds were

winnowed, sorted, washed and drained. After draining the unfermented samples (sliced sweet

potato and pigeon pea) wet milled, sieved, sedimented for 24 hours, drained, sun dried for 3 days

39
and dry milled into powder, sieved and packaged in airtight container for further analysis (Fig.

2).

Sweet potato tuber (70%) Pigeon pea (30%)

Washing Winnowing

Peeling Peeling

Washing Washing

Slicing Soaking (72 hrs at room temp.)

Draining

Re-washing

Wet milling

Sieving

Sedimentation (24 hrs)

Draining

Caking

Sun drying (72 hrs)

Dry milling

Packaging

Fermented sweet potato-pigeon pea flour

40
Figure 1: Preparation of Fermented sweet potato-pigeon pea flour
Sweet potato tuber (70%) Pigeon pea (30%)

Washing Winnowing

Peeling Peeling

Washing Washing

Slicing Draining

Wet milling

Sieving

Sedimentation (24 hrs)

Draining

Caking

Sun drying (72 hrs)

Dry milling

Packaging

Unfermented sweet potato-pigeon pea flour

Figure 2: Preparation of unfermented sweet potato-pigeon pea flour

41
3.2.3 Preparation of blends for weaning food

Sweet potato flour was blended with pigeon pea flour, in different combination to obtain

Samples (2) samples as shown in Table 1.

FSPWF = Fermented Sweetpotato (70%) + Pigeon pea (30%) Weaning Food

USPWF = Unfermented Sweetpotato (70%) + Pigeon pea (30%) Weaning Food

42
Table 3.1: Flour Blends Formulations

Samples Sweet potato flour Pigeon pea flour

FSPWF 70 30

USPWF 70 30

Keys: FSPWF = Fermented Sweetpotato-Pigeon pea Weaning Food

USPWF = Unfermented Sweetpotato-Pigeon pea Weaning Food

43
3.3 Microbial Evaluation of weaning food

3.3.1 Preparation of Media

The three media (Nutrient Agar, (NA); MacConkey agar, (MAC); Potato Dextrose Agar,

(PDA)) used were prepared according to the manufactures’ procedures (Titan Biotech Ltd.

BHIWADI-301019, Rajasthan, India).

3.3.2 Preparation of Nutrient Agar (NA)

Twenty eight grams (28.0g) of nutrient agar powder was dissolved in 1000 ml of distilled

water. The medium was gently heated to dissolve and completely sterilize by autoclaving at

15psi (121°C) for 15minutes.The autoclaved media was allowed to cool before pouring 15ml

each onto sterile Petri dishes and allowed to gel.

3.3.3 Preparation of MacConkey agar (MAC) Agar

This was done by suspending 47grams of the powder in 1000ml of distilled water. The

suspended was mixed very well and heated with frequent agitation to dissolve the powder

completely. The suspended media was sterilized by autoclaving at 121°C and 15psi, for

15minutes. The autoclaved media was allowed to cool before pouring 15ml each onto sterile

Petri dishes and allowed to gel.

3.3.4 Preparation of Potato Dextrose Agar (PDA)

This was done by suspending 39g of the media in 1000ml of distilled water. The

suspended was mixed very well and heated with frequent agitation to dissolve the powder

completely. The suspended media was sterilized by autoclaving at 121°C and 15psi, for 15

44
minutes. The autoclaved media was allowed to cool before pouring 15ml each onto sterile Petri

dishes and allowed to gel.

3.3.5 Microbiological Analysis

One milliliter of each sampled was put in 9ml of sterile distilled water in sterile test

tubes, shaken and then serially diluted. From the appropriate dilution, 0.1ml was inoculated

separately on to MacConkey agar, Nutrient Agar and Potato Dextrose Agar plates and spread

evenly using sterile bent glass rod. Each experiment was carried out in duplicates to get a mean

standard value of the colony forming units (cfu/ml) on the plates. The inoculated MacConkey

agar, Nutrient agar and Potato Dextrose agar plates were incubated at 30ºC and 35ºC for 24 and

48 hours respectively. After the period of incubation, the colonies on the plates were counted and

recorded as colony forming unit per milliliter (cfu/ml) and coliform respectively (Harrigan and

McCance, 2006). Each of the bacterial colonies on the agar plates was sub-cultured and the pure

culture obtained. Isolates were identified by carrying out tests which include Gram staining,

spore staining and biochemical tests such as catalase, coagulase, oxidase, citrate utilization,

indole, methyl red, urease, VogesProskauer and sugar fermentation (Onyeagba, 2004;

Cheesbrough, 2006).

The bacteria isolates were characterized using microscopic techniques and biochemical

tests. The identities of the isolates were determined by comparing their characteristics with those

of known taxa as described by Bergey’s manual of Determinative Bacteriology (Holt et al.,

2004) and Cheesbrough, (2006). The cultures used for biochemical test were between 18 to 24

hours old.

45
 Fungal identification and enumeration was based on their colony elevation, colour,

texture, shape and arrangement of conidia (spherical or elliptical, unicellular or multicellular),

branched or unbranched mycelia, presence or absence of cross walls (whether septate or non-

septate) and others. They were enumerated by reference to illustrated manual on identification of

some seed borne fungi (Kulwant et al., 2001) and illustrated genera of imperfect fungi (Barnet

and Hunter, 2002).

46
 CHAPTER FOUR

4.0 RESULTS AND DISCUSSION

4.1 Results

Table 4.1: Microbial analysis of fermented and unfermented sweet potato-pigeon pea

weaning food

Samples Yeast & Mould count Total viable count Coliform count

FPSWF 2 × 10-5 5 × 10-5 0

UPSWF 6 × 10-5 5 × 10-5 0

Keys: FPSWF = 70% Fermented sweet potato + 30% pigeon pea weaning food

UPSWF = 70% Unfermented sweet potato + 30% pigeon pea weaning food

47
4.2 Discussion

Microbial analysis of fermented and unfermented sweet potato and pigeon pea

(weaning food)

Microorganisms play significant role in the determination of shelf life of food products.

They are usually responsible for spoilage of many food items. A high aerobic plate count (APC)

could indicate the presence mixed population of microorganisms, which may consist of spoilage

types. The microbial count of weaning food produced from incorporation of pigeon pea and

sweet potato, fermented and unfermented samples is shown in the Table 4.1. The total viable

count for the unfermented weaning sample (UPSWF) sample was the same with the fermented

samples (FPSWF) with the value of 5×10-5 Cfu/g as shown in the table above. Sample (FPSWF)

had the lowest microbial load from the result obtained in the analysis. This result differed from

that obtained by Samadrita et al. (2012) who compared the microbial load of different okara

samples based on processing (drying methods). The results from the study suggest that the

fermented weaning food should be used in further applications as soon as possible since

prolonged storage may negatively impact on their proximate composition (the feature that made

them to be highly valued for food and feed formulations in the first place). There was no

significant difference (p<0.05) between both samples in the total viable count.

From the result obtained in the study, there was no coliform count in the samples which

could be because the processing of the samples were carried out in a clean environment, they

were dried in an oven drier and clean containers were used to package the blends of the samples

after processing.

48
 In the yeast and mould count, sample (UPSWF) was identified to have the highest growth

of 6.0 × 10-5 Sfu/g which is low to cause harm to consumers whereas there was low detection of

microorganisms in FPSWF 2.0 × 10-5. The acceptable level for mould in breakfast cereals is 10

Sfu/g whereas the level which when exceeded in one or more samples would cause the lot to be

rejected as this indicate potential health hazard or imminent spoilage is 103 Sfu/g (FDA, 2013).

The acceptable level for mould in flour is 103 Sfu/g whereas the level which when exceeded in

one or more samples would cause the lot to be rejected as this indicate potential health hazard or

imminent spoilage is 105 Sfu/g (FDA, 2013). There was significant difference (p<0.05) between

the samples.

49
 CHAPTER FIVE

5.0 CONCLUSION AND RECOMMENDATIONS

5.1 Conclusion

This study has shown that microbiological qualities of fermented and unfermented

weaning were not significantly affected by incorporation of pigeon pea and sweet potato.

Conclusively, pigeon pea and sweet potato can be used to produce a weaning food of high

nutritive and enhance its microbial properties. The low microbial count (below the acceptable

safe level) was reported.

5.2 Recommendation

Limits of microbial counts have been recommended in most foods to keep them safe for

consumption. Thus, there is need to properly store product with packaging best suitable for it and

in the best environment in order to prevent contamination and more microbes from gaining

entrance. It is recommended that further study should be carried out on the health benefits of

consuming fermented and unfermented weaning food with incorporation of pigeon pea and sweet

potato. It is also recommended that consumers should be introduced weaning food as it is not

popularly known, especially in the western part of Nigeria where this work was carried out. It is

recommended that further study be carried out on the following: Nutritional (proximate)

composition of other fermented and unfermented products; Shelf-stability studies on weaning

(fermented and unfermented); Isolation and characterization of the fermenting microflora in the

products; Feeding trials using bioassay.

50
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