ABSTRACT

THE STUDY EXAMINED THE VARIOUS FUGAL PATHOGENS ASSOCIATED WITH TOMATOES
SPOILAGE AS WELL AS THEIR PRELIMINARY TAXONOMIC GROUPING UP TO GENUS LEVEL. 30
SAMPLES OF THE SPOILED TOMATOES WERE COLLECTED FROM AFAHA NSITR.
MORPHOLOGICAL STUDIES REVEAL THE FUNGI MEMBERS WHICH INCLUDE MEMBERS OF THE
GENUS ASPERGILLUS, CLADOSPORIUM, FUSARIUM. SPECIES OF FUSARIUM WERE SEEN TO BE
THE MOST OCCURRING IN ALL SAMPLES. STATISTICAL ANALYSIS ALSO CONCURRED TO THE FACT
THAT FUNGI ARE RESPONSIBLE FOR SPOILAGE OF TOMATOES PURCHASED IN AFAHA NSIT. IT
IMPLIES THAT FUNGI ARE GREATLY RESPONSIBLE FOR TOMATO SPOILAGE. PROPER HANDLING,
TRANSPORTATION AND THOUGH WASHING WITH CLEAN OR CHLORINATED WATER WILL GO A
LONG WAY IN REDUCING THE RISK OF TOMATOES SPOILAGE BY FUNGI.

CHAPTER ONE
1.0 Introduction

Vegetables constitute commercially and nutritionally important indispensable food commodity.

Vegetable naturally play a vital role in human nutrition by supplying the necessary growth
factors such as vitamins and essential minerals in human daily diet and that can help to keep a
good and normal health. Vegetables are widely distributed in nature. One of the limiting factors
that influence the fruit economic value is the relatively short shelf-life period caused by
pathogen’s attack (Droby, 2006: Zhu, 2006).

Tomato (Lycopersiconesculentum) pronounced Toh-MAH-to is the most important vegetable

worldwide. Tomato is an annual plant which belongs to the Solanaceaefamily which includes
other well-known species such as Potato, Tobacco, Pepper and Egg plants (aubergine) and can
reach a height of over two meters. It’s a berry fruit, tomato is grown mainly in soil (Frazier and
Westhoff,2004).

Tomato is essential mainly for its dietary needs and can be consumed in diverse ways. It can be
cooked as vegetable, as an ingredient in many dishes and sauces; in the making of stew, fruit
juices and can be eaten raw in salads(Masefield et al., 2002).

The modern day tomatowhich is a very popular fruitoriginated from South-Western

Hemisphere, now the Peru-Euador area(Rick,2001).From here,it was distributed to many parts
of tropical America. It possibly arrived the West African Ports via Portuguese traders or the
Sudan (Tindall, 2004).

Tomato in West Africa is grown in gardens and irrigation schemes. In Nigeria, most tomatoes
are grown in the northern parts of the country(Erinle,2007) and there is no record of any
systematic or organized traditional storage method for vegetables and fruits. They are usually
sold immediately after harvesting. They are packed in baskets, cardboard boxes, or wooden
crates ready for transportation to the markets.

Tomato is the most perishable vegetable during handling, transportation and storage. This is
because tomato contains large amount of water which makes them susceptible to spoilage by
the action of microorganisms such as fungi, bacteria and protozoans (Ross, 1975). Estimates
have shown that about one third of the produce is lost before reaching the consumer (Erinle,
2007), this loss has been attributed to a number of factors which include physical (mechanical
breakage, bruises), physiological and also damages caused by pathological agents (Thompson
and Kelly, 2000), market value of the tomato are mainly reduced by this factors.

Watt and Merrill, (2000) defined spoiled food as that which have been damaged or injured so as
to render it undesirable for human consumption. Various activities may result in food spoilage:
insect activities,causing physical injuries for example, bruisingand enzyme activity by
microorganism causes change in colour, taste, smell, texture and quality of the food.

Pauland Sarisbury, (1981) also referred to spoilage as any change in the condition of food in
which the food becomes less palatable or even toxic: these changes may be accompanied by
alteration in taste, smell, appearance or texture.

The spoilage of food by microorganisms should not be viewed as a sinister plot on the part of
the microbes deliberately to destroy foods but as a normal function of these organisms in the
total ecology of all living organisms (Watt and Merrill, 2000). It has also been estimated by Watt
and Merrill (2000) that 20% of all fruits and vegetables harvested for human consumption are
lost through microbial spoilage causing one or more of 250 market diseases. Spoilage of fresh
tomato usually occurs during storage and transit and also while waiting to be processed.

Frazier and Westhoff,(2004) reported that fruits and vegetables after picking continue to
respire and as a result heat is produced, this heat is known as ‘vital heat’, the amount which
varies with particular amount of fruits and vegetables.

Vegetables that respire more thereby generating most heat are those that perish/spoil most
rapidly (Wheeler, 2005). The resulting respiration of the fruits in addition to the normal ripening
process complicates an independent discussion on microbiological spoilage of fruits in general
and tomato in particular.

Fusarium oxysporium f.sp.lycoperisici andVerticilliumalbo-atrumare two fungi that cause similar

wilt disease on tomato.(Nesmith etal.,1983). Fusarium is more common in replant fields and
gardens while Verticilliumhas been destructive on greenhouse tomatoes and in commercial
stake tomatoes. Verticillium wilt is more of a problem at cooler temperature while Fusarium
wilt is considered a warm temperature disease. Both are favoured by wet conditions.

According to Nesmithetal., (1983), both fungi are common inhabitants of Kentucky soils. These
fungi attack the plant through the roots and grow up through the water-conducting vessels.
FusariumandVerticilliummay be introduced to soils in several ways: Old crop residues,
transplants, wind, water,implement-borne soils or mulches. These fungi become established
readily in most soils and can remain in the soil for years. When susceptible tomatoes are
planted, in infected soil, their roots are also subjected to attack by these fungi. Both diseases
are much more serious when accompanied by root knot nematode. These problems can be
controlled by using resistant tomato varieties and fumigating or stream sterilizing greenhouse
soils.

1.1 Statement of the Problem

Tomatoes are an attractive cash crop for small scale farmers and provide potential source of
employment to many Nigerians. Without a doubt, the tomato fruits have been marketed
freshly picked from the field and is the best selling fresh market vegetable crop.

Despite the human need of tomato, damage as a result of spoilage caused by micro-organisms
has been of serious concern. The isolation and characterization of fungi associated with spoiled
tomatoes.

Microbial decay is one of the main factors that determine losses and compromises the quality
of the produce. The extent of the losses especially through microbial decay has not been
quantified in most areas and where this has been quantified the results are short lived. Both the
biological and physical damages during the harvest and transportation phases, coupled with
large amount of water and soft endocarp makes tomatoes more susceptible to spoilage by
fungi.

Therefore, in order to provide literature on this area, few studies on fungi associated with
tomato spoilage are available in Nigeria. This then motivated the researcher to investigate into
isolation and characterization of fungi associated with spoiled tomatoes.
1.2 BACKGROUND OF THE STUDY

Tomatoes (Solanum lycopersicum) is perishable vegetable widely cultivated and consumed

worldwide (Agrios, 2005; Valadez et al., 2012). Originated in West South America (kimura and
Sinha, 2008).Tomatoes is an annual plant which belongs to the Solanaceae family. Tomatoes
are essential mainly for its dietary needs and can be consumed in diverse ways. It can be
cooked as vegetable, as ingredient in many dishes and sauces; in making of stew, fruit and can
be eaten raw in salad. It is rich in nutrients, vitamin, dietary fibres and phytochemical (Bradley,
2003; Freeman and Reimers, 2011; Mariga, 2012).They are good source of natural antioxidant
which include carotenoids, vitamin phenolic compound Flavonoid which have shown to
eliminate free radicals (Sravanthi and Grangadhar, 2005; Pitchaon et al., 2007). Tomatoes has a
much lower sugar content than other fruits and is therefore not as sweet and their low pH
values make them particularly desirable to fungal decay. Pathogenic fungi, such as Aspergillus,
Fusarium, Mucor, Penicillium, Rhizopus, cladosporium species have been implicated in some
crop spoilage (Beuchat, 1995). Fungi contamination in many Agricultural Product including
tomatoes starts in the fields.

Both biological and physical damage during the harvest and transportation phases, coupled
with large amount of water and soft endocarp makes tomatoes more susceptible to spoilage by
fungi (Asan and Ekmeiki, 2002). Spoilage of tomatoes are those adverse changes in the quality
of tomatoes that are brought about by the action of predominantly biological and physical
factors. These may be changes in taste, smell, appearance or texture of the fruit. (Ghosh, 2009)
reported that fungi were the source of spoilage of most of the tomatoes sample accessed than
bacteria. This fruit are usually displayed on benches and in baskets for prospective customers in
the open markets until sold thereby exposing them to further microbial infection beside those
associated with these whole fruit surface and those from adjacent infected fruits (Baijewu et al,
2007). Few studies on fungi associated with tomato spoilage are available in Nigeria (Wogu and
Ofuase, 2014; Wokoma, 2018; Eni et al., 2010; Bashir et al., 2016). Eating or drinking
contaminated foods or drinks can cause food borne disease (Adebajo et al; 2012). Fungal
spoilage of tomatoes has been recognized as a source of potential health hazard to humans and
animals, they produce mycotoxins which are capable of causing mycotoxicoses in man following
ingestion or inhalation (Baker, 2006).The mycotoxins are not limited to their areas of infections.
Since tomatoes contain large amount of fluid, these mycotoxins diffuse rapidly throughout
them, contaminating all parts and making the fruit unfit for consumption. The importance of
tomatoes in the food industry and its nutritional benefits cannot be overemphasized. Routine
microbiological examination of tomatoes is very crucial as it contributes to a large extent to
economic development.

1.3 Justification

Since microorganisms have been identified with the major cause of spoilage in fruits and
vegetables due to their high water content, proper isolation and characterization of these
organisms in tomatoes will greatly reduce the spoilage of this perishable fruit and as such
producers and consumers will be able to protect their vegetables (tomato) and also identify
spoiled tomatoes that have been attacked by fungi.

Works have been done on the isolation and identification of bacteria and fungi on fruits(Okoli,
1998). Little or no work has been done on the isolation and characterization fungi on spoiled
tomatoes in Gwagwalada.Therefore, the major organisms responsible for field and market
diseases will be majorly identified and checked.

1.4 Aim And Objectives

This research work is aimed at identifying the various fungi organisms associated with the
spoilage of fresh tomatoes in Gwagwalada with the view of mapping out strategies to curtail
these fungal attacks and also to determine the effects of the isolated fungi on the spoiled
tomatoes.

Objectives

1. To isolate of fungi organism from spoilt tomatoes in Gwagwalada.

2. To characterize and identify of the fungi in spoilt tomatoes.
3. To determine the prevalence and pathogenicity of fungal organisms.
 4. To create public health awareness about the preponderance of fungal organisms.

1.5 Significance of the study

This study provides important information on the current status on isolation and
characterization of fungi species associated with spoiled tomatoes in Nigeria and their
management using crude plant extracts. The outputs of this study will be important in reducing
tomato post-harvest losses by improving the efficiency of post-harvest handling due to better
post-harvest management strategies.

1.6 Research Questions

The following research questions are stated to guide the study:

(i) What are the fungi species associated with spoiled tomatoes in Gashua?

(iii) What are fungi species associated with spoiled tomatoes in Gashua?

1.7 Limitation Of Study

Researches have been conducted to ascertain the fact that various microorganisms are
responsible for the spoilage of tomatoes. Organisms include Bacteria, for
example,Pseudomonas and Xanthomonasspeciesas stated by Watt and Merrill (2000), Fungi,
for example, Alternaria, Colletotrichum ,Fusariumand Penicillium species as stated by Barksdale
(2001) and other Meioidogyne groups of nematode.

This present study places emphasis on the spoilage of tomatoes in Gwagwalada Area Council
caused by pathogenic fungi species.

1.8 Hypothesis

H0; Fungi are not responsible in the spoilage of tomatoes.

HI; Fungi are responsible in the spoilage of tomatoes.

1.9 Scope of the Study

Researches have been conducted to ascertain the fact that various microorganisms are
responsible for the spoilage of tomatoes. Organisms include Bacteria, for example,
Pseudomonas and Xanthomon as species as stated by Watt and Merrill (2000). This present
study places emphasis on isolation characterization of fungi species associated with spoiled
tomatoes in Nigeria.

1.10 Limitation of the Study

Despite that this research work was to highlight the microorganism association with spoilage of
tomato, it was free with a lot of hindrances which include:

Time Constant: The research was constrained by the time limit, it posed a treat to the
successful coverage intended in the course of this study. We also have a problem of managing
time because we needed time to attend lectures which we have the time to do our practical.

Cost Factor: The presence of economic hardship in our country posed to a great extent a
problem in proving the whole material, information (and transportation) needed other
statistical data procured was also limited by cost factor.

Unavailability of research material.

1.11 operational definition of terms

Isolation: solitude, isolation, seclusion mean the state of one who is alone. solitude may imply a
condition of being apart from all human beings or of being cut off by wish or circumstances
from one's usual associates. a few quiet hours of solitude. isolation stresses detachment from
others often involuntarily.

Characterization: Characterization tells the audience what the personality of the character is.
Example: “The patient boy and quiet girl were both well mannered and did not disobey their
mother.” Explanation: The author is directly telling the audience the personality of these two
children.
Fungi: any member of a kingdom of organisms (Fungi) that lack chlorophyll, leaves, true stems,
and roots, reproduce by spores, and live as saprotrophs or parasites. The group includes
moulds, mildews, rusts, yeasts, and mushrooms.

Spoiled: having the character or disposition harmed by pampering or oversolicitous attention

Tomatoes: is the edible berry of the plant Solanum lycopersicum, commonly known as the
tomato plant. The species originated in western South America, Mexico, and Central America.
The Nahuatl word tomatl gave rise to the Spanish word tomate, from which the English word
tomato derived. Its domestication and use as a cultivated food may have originated with the
indigenous peoples of Mexico

CHAPTER TWO

Review of related literature

2.1 Brief history of Tomatoes Cultivation

Tomato (Lycopersicon esculentum Mill.) is one of the most popular and widely grown plants in
the world as well as in Africa (Osemwegie, et al., 2010). It is the second most important
vegetable worldwide, in terms of the amount of vitamins and minerals it contributes to the diet
(Enrique & Eduardo, 2006).

Genetic evidence shows that the progenitors of tomatoes were herbaceous green plants with
small green fruit and a center of diversity in the high lands of Peru (Smith 1994).
The Spanish explorer, Cortes may have been the first to transfer the small yellow tomato fruits
into Europe after he had captured the Aztec city of Tenochtitlan, now Mexico City in 1521;
although Christopher Columbus, a Genoese working for the Spanish monarchy may have taken
them back as early as 1493.

The earliest discussion of tomato in European literature appeared in an herbal written in 1541
by Pietro Andrea Mathiolian Halian, an Italian physician and botanist, who named it pomod’oro,
or ‘gold apple’ (Smith 1994). One of the earliest cultivators was John Gerard, an Herbal-
Surgeon. Smith (1994) reiterates that Gerard’s Herbal, published in 1597, and largely
plagiarized from continental sources, is indeed one of the earliest discussions of tomato in
England.

Similarly, Tindall (1988) remarked that the Portuguese trader and freed slaves from West Indies
introduced this plant into West Africa Countries.

2.2 Description and Scientific classification of tomatoes

Scientific classification

Kingdom: Plantae

Clade: Tracheophytes

Clade: Angiosperms

Clade : Eudicots

Clade: Asterids

Order: Solanales

Family: Solanaceae

Genus: Solanum

Species: S. lycopersicum
Binomial name Solanum lycopersicum L.

Tomatoes are a significant source of umami flavor. They are consumed in diverse ways: raw or
cooked, and in many dishes, sauces, salads, and drinks. While tomatoes are fruits—botanically
classified as berries—they are commonly used culinarily as a vegetable ingredient or side dish.

Numerous varieties of the tomato plant are widely grown in temperate climates across the
world, with greenhouses allowing for the production of tomatoes throughout all seasons of the
year. Tomato plants typically grow to 1–3 meters (3–10 ft) in height. They are vines that have a
weak stem that sprawls and typically needs support. Indeterminate tomato plants are
perennials in their native habitat, but are cultivated as annuals. (Determinate, or bush, plants
are annuals that stop growing at a certain height and produce a crop all at once.) The size of the
tomato varies according to the cultivar, with a range of 1–10 cm (1⁄2–4 in) in width.

Toxicity

The leaves, stem, and green unripe fruit of the tomato plant contain small amounts of the
alkaloid tomatine, whose effect on humans has not been studied. They also contain small
amounts of solanine, a toxic alkaloid found in potato leaves and other plants in the nightshade
family. However, solanine concentrations in foliage and green fruit are generally too small to be
dangerous unless large amounts are consumed—for example, as greens.

Small amounts of tomato foliage are sometimes used for flavoring without ill effect, and the
green fruit of unripe red tomato varieties is sometimes used for cooking, particularly as fried
green tomatoes. There are also tomato varieties with fully ripe fruit that is still green.
Compared to potatoes, the amount of solanine in unripe green or fully ripe tomatoes is low.
However, even in the case of potatoes, while solanine poisoning resulting from dosages several
times the normal human consumption has been demonstrated, actual cases of poisoning from
excessive consumption of potatoes are rare.

Tomato plants can be toxic to dogs if they eat large amounts of the fruit, or chew plant
material.
Salmonella

Tomatoes were linked to seven Salmonella outbreaks between 1990 and 2005, and may have
been the cause of a salmonellosis outbreak causing 172 illnesses in 18 US states in 2006. The
2008 United States salmonellosis outbreak caused the temporary removal of tomatoes from
stores and restaurants across the United States and parts of Canada, although other foods,
including jalapeño and serrano peppers, may have been involved.

Uses

Though it is botanically a berry, a subset of fruit, the tomato is a vegetable for culinary purposes
because of its savoury flavour. One study has shown that the inner pulp has a deeper umami
taste than the outer flesh.

Although tomatoes originated in the Americas, the tomato is now grown and eaten around the
world. It is used in diverse ways, including raw in salads or in slices, stewed, incorporated into a
wide variety of dishes, or processed into ketchup or tomato soup. Unripe green tomatoes can
also be breaded and fried, used to make salsa, or pickled. Tomato juice is sold as a drink, and is
used in cocktails such as the Bloody Mary.

Tomatoes have become extensively used in Mediterranean cuisine as a key ingredient in pizza
and many pasta sauces. Tomatoes are also used in Spanish and Catalan dishes, such as
gazpacho and pa amb tomàquet.

Storage

Tomatoes keep best unwashed at room temperature and out of direct sunlight. It is not
recommended to refrigerate them as this can harm the flavour.[105] Tomatoes stored cold
tend to lose their flavour permanently. Storing stem down can prolong shelf life, as it may keep
from rotting too quickly.Tomatoes that are not yet ripe can be kept in a paper bag till ripening.

Tomatoes are easy to preserve whole, in pieces, as tomato sauce or paste by home canning.
They are acidic enough to process in a water bath rather than a pressure cooker as most
vegetables require. The fruit is also preserved by drying, often in the sun, and sold either in
bags or in jars with oil.

2.3 Temperature requirement for tomatoes

Tomato is a warm season crop. The best fruit colour and quality is obtained at a temperature
range of 21-24°C. Temperatures above 32o C adversely affects the fruit set and development.
The plants cannot withstand frost and high humidity. It requires a low to medium rainfall. Bright
sunshine at the time of fruit set helps to develop dark red coloured fruits. Temperature below
10 oC adversely affects plant tissues thereby slowing down physiological activities.

Tomatoes love the sunshine. A position in full sun (that means an average of at least eight
hours a day) gives the best results in most areas, though if you're in a hot climate you can get
away with dappled shade. Under a high temperature condition, the fertility rate of tomato
flowers is greatly reduced, leading to flower drop and reduced fruit setting.

Actually, tomatoes like warm weather, between 65 and 85 degrees. When temperatures soar
past 95, tomatoes stop growing. In that kind of heat, their flowers fail to pollinate and instead
they dry up and drop off, putting a pause on the production of new fruit.

The minimum temperature is around 10˚C with the maximum being 34˚C. The optimum
temperature is between 26 and 29˚C. For the optimal development of the tomato plant, a
relative humidity between 65% and 85% is required. Higher relative humidity levels negatively
influence pollen release and distribution on the stigma.

2.4 Soil type and pH range of tomato

Tomatoes are one of the most popular crops among home gardeners, and with good reason:
not only are tomatoes delicious and versatile, they are relatively easy to grow and return high
value for the space they occupy. No store-bought tomato can compare with the flavor of a vine-
ripened tomato picked from the garden at its peak of ripeness.

Tomatoes are one of the most popular vegetables grown in home gardens and on farms. They
are known for their versatility in cooking and their delicious taste. However, to grow healthy
and bountiful tomatoes, it is important to understand the role of soil pH. In this article, we will
be discussing the importance of soil pH for tomatoes, and how it affects plant growth.

Brief Overview of the Importance of Soil pH for Tomatoes

Soil pH is a measure of the acidity or alkalinity of the soil. It is measured on a scale of 0 to 14,
with 7 being neutral. A pH level below 7 is considered acidic, and above 7 is alkaline. The pH
level of the soil can have a significant impact on the growth and health of plants. For tomatoes,
the ideal pH level is between 6.0 and 7.0. If the pH level is too high or too low, it can negatively
affect the availability of essential nutrients for the plant. This can lead to stunted growth,
reduced yield, and even plant death.

A perennial plant in its native tropics, tomato belongs to the nightshade family (Solanacae) and
is native to Central and South America. The tomato appeared in European cuisine in the 16th
century, although it did not become popular there until the 18th century because of the
common belief that it was poisonous.

Soil Requirements

Tomato plants will grow well in well-drained sites that receive full sun for most of the day. The
soil pH should be slightly acidic (6.2 to 6.8). Excess nitrogen can result in plants with lush,
vigorous foliage but little fruit production. Although it is best to determine lime and fertilizer
needs from the results of a soil test, a rule of thumb for gardeners lacking test data is to apply
2½ pounds of a complete fertilizer such as 10-10-10 (or the equivalent) per 100 square feet of
garden area. Work the fertilizer into the soil about 2 weeks before planting.

To put it simply: tomatoes get their flavour from the soil in which they grow, just like wine.
Flavoursome tomatoes need loamy soil that are rich in humus. Good quality peat-based potting
compost is just the ticket. Tomato can be grown on a wide range of soils from sandy to heavy
clay. However, well-drained, sandy or red loam soils rich in organic matter with a pH range of
6.0-7.0 are considered as ideal.
In the case of tomatoes, peppers, and a few other vegetables, it contributes to a problem called
blossom end rot. Vegetables need a soil pH between 6.2 and 6.8 (herbs prefer a pH near 7.0,
which is called neutral). When it comes to growing tomatoes, the ideal soil pH is between 6.0
and 7.0. This range is considered to be slightly acidic to neutral and is the optimal range for the
availability of essential nutrients for the plant. At a pH level of 6.0 to 7.0, the soil can provide
the right balance of nutrients for the tomato plant to thrive. The roots can absorb essential
nutrients such as nitrogen, phosphorus, and potassium, allowing the plant to grow strong and
healthy. Additionally, this pH range also promotes the growth of beneficial microorganisms in
the soil, which aid in the breakdown of organic matter and the release of nutrients for the plant
to use.

2.5 Tomato diseases

Soil-borne fungal diseases can be a major problem of tomatoes. There are three steps to
understanding and managing tomato diseases in the home garden. The first step is to
understand the disease cycle of a typical fungus. The second is to recognize symptoms of
important fungal diseases of tomato, and the third is to apply good cultural practices to help
minimize the damage caused by these diseases.

Simply, fungi live and obtain their nourishment from infected host tissue. Fungi reproduce by
spores, tiny microscopic bodies, which are spread by wind, water, or other mechanical means
to a new host. On the host, spores germinate and infect healthy plant tissue causing symptoms
including leaf spots, rots, and wilts that lead to premature defoliation and reduced tomato
yields. Development and spread of fungi in the home garden is determined by rainfall, relative
humidity, free moisture, and temperature.

Some of the most common fungal diseases that infect tomatoes grown in the home garden
include Anthracnose fruit rot, Early blight, Septoria leaf spot, Late blight, and Buckeye rot all
which produce distinct symptoms making them easily diagnosable by the home gardener.

Anthracnose Fruit Rot

Anthracnose fruit rot is a soil-borne disease that affects ripe tomato fruit. Infections go
unnoticed on green fruit and as fruit ripens depressed circular water-soaked spots appear on
red fruit. These spots may slowly enlarge to about 1/4-inch in diameter and produce black
fungal structures (microsclerotia) in the center of the lesion just below the skin surface.
Microsclerotia can overwinter in the soil and serve as a source of inoculum for the next growing
season.

Early Blight

Early blight can infect tomato foliage and fruit. On tomato foliage, Early blight first appears as
circular irregular black or brown spots on the older leaves of the plant. As these lesions enlarge
a series of dark concentric rings develop in the center of the spot creating a distinct target
pattern. Over time the tissue surrounding the early blight lesions can yellow and cause the
leaves to drop. Severe infestations of this disease can cause 100% defoliation of the plant.

Early blight can infect the fruit through the calyx or stem attachment in the immature green or
red stage and can produce distinct target-like lesions similar to foliar infection. Defoliation
caused by early blight can reduce fruit yield and can leave the fruit open to sunscald injury.

Septoria Leaf Spot

Septoria leaf spot is a soil-borne fungal disease that only infects tomato leaves and stems. The
spots enlarge to 1/8-inch in diameter and are distinguished by a dark brown edge with a white
or gray center. As the disease develops and more leaf spots develop, the areas surrounding
spots will turn yellow causing leaves to wither and die. The disease spreads from the older
leaves upward and can totally defoliate a plant in a short period of time. Defoliation can
severely inhibit yield and lead to sunscald injury and fruit cracking. If tomato plants become
infected with Septoria leaf spot early in the growing season, plants can become 100%
defoliated before fruit set.

Late Blight
Late blight is the disease historically associated with potatoes and the Irish potato famine of the
mid-1800's. The Late blight fungus can also infect tomato plantings. The disease will first appear
as greasygrayish indefinite patches on older leaves and stems. These spots enlarge in moist
weather and may produce white fuzzy growth on the underside of infected leaves. The fungus
will also attack fruit causing a dark, greasy colored lesion with a slightly sunken, rough surface
on green fruit. These lesions may enlarge turning the whole fruit brownish-black. Infected fruit
often remain firm. Severe infestations can cause the foliage to brown and shrivel.

Buckeye Rot

Buckeye rot typically affects immature and mature green fruit that lay on the soil surface. On
green fruit, buckeye rot will produce distinct brownish-black lesions that have a definite
concentric or target-like appearance. In moist conditions, white fluffy fungal growth will
develop on infected fruit. Over time, infected fruit will become soft and mushy.

2.5.1 Field diseases

Soil borne diseases are those plant diseases caused by pathogens who inoculate the host by
way of the soil (as opposed to the air or water). Unhealthy soils can have issues with high levels
of disease incidence and pests. Common soil borne diseases include damping-off, root rot and
vascular wilt; and can exhibit symptoms such as tissue discoloration, wilting of foliage, root
decay and sudden death.2 Soil-borne diseases can significantly reduce yields for many crops,
and can decimate the agricultural sectors of large areas if not managed carefully.

Pathogens, the biological agents responsible for soil borne diseases, are drawn from several
taxonomic groups. The largest group are the fungi, but plant diseases can also be caused by
bacteria, protozoa, viruses and nematodes. Under certain conditions, many of these disease
causing organisms already exist in the soil in a non-pathogenic form. Other invasive organisms
are only able to survive in the soil for a short amount of time after being introduced. In both
cases, changes in soil factors such as pH, moisture, temperature and nutrient level can
precipitate a disease outbreak. These changes can be a result of a shift in agricultural practices,
introduction of new substances to the soil, or introduction of new crops.
Management strategies such as ensuring well-drained, well-aerated fields; selection of disease-
resistant plant varieties; and application of compounds such as copper sulfate can aid greatly in
reducing soilborne disease incidence.

2.5.2 Transit and market disease of Tomato

After the tomatoes are harvested, they are transported to a processing plant. This is typically
done using large trucks, which are specially designed to transport perishable goods like fresh
tomatoes. During transportation, the tomatoes must be kept at the appropriate temperature to
ensure their freshness and quality.

In transporting tomatoes over long distances the aim is to have them reach the consumers
without any deterioration in the quality. The tomatoes must be firm, of a fine colour both inside
and outside, and they must have a good taste. The optimum storage temperature in the
warehouse and during transport is 12°C and the humidity 85–90% relative humidity.

Tomatoes are packaged in open and closed fruit crates (tomato crates), tubs, cartons, trays and
jointed boxes. Sometimes they are carefully arranged, sometimes randomly bulk-packed.
Broken, damaged and damp packages must be rejected. Refrigerated container with fresh air
supply or controlled atmosphere.

Tomatoes are produced commercially both in the field and in enclosed structures (high tunnels
and greenhouses). They are also a very popular garden plant among homeowners. Tomato
production, whether for commercial or personal use, is not always an easy task. A variety of
disorders, insects, diseases, and pests may cause problems during any given growing season
and may damage a crop, leading to reduced or poor-quality yields.

Some of the most common fungal diseases that infect tomatoes grown in the home garden
include Anthracnose fruit rot, Early blight, Septoria leaf spot, Late blight, and Buckeye rot all
which produce distinct symptoms making them easily diagnosable by the home gardener.
A number of disease-causing organisms (pathogens) can infect tomatoes and cause disease.
Many of the most common diseases that affect tomatoes in Mississippi are caused by various
fungi, bacteria, and viruses. A few fungus-like organisms (oomycetes) commonly called water
molds also cause troublesome diseases in Nigeria. Many diseases of tomatoes occur both in the
field and in enclosed structures; however, because of the unique nature and environmental
conditions present in enclosed structures, some diseases are more common in these structures,
whereas others are more common in the field. Additionally, some disease management
methods for tomatoes grown in greenhouses are unique to those structures since the
environment in these structures can be manipulated.

2.6 Factors that influence spoilage of fruits and vegetables

Food spoilage and deterioration is no accident. It is a naturally occurring process. Some of the
primary culprits are air, moisture, light, temperature, and microbial growth. Most fruits and
vegetables spoil easily because of damage caused by microorganisms. Microorganisms such as
bacteria, yeast and molds need water and nutrients for growth, energy and reproduction.

Factors that affect food spoilage include:

 Microorganisms
 Enzymes
 Air
 Light
 Insects, Rodents, Parasites and Other Creatures
 Physical Damage
 Temperature
 Time

Microorganisms
Many types of microorganisms can cause food problems. The microorganisms that can cause
food-borne illness are called pathogenic microorganisms. These microorganisms grow best at
room temperatures (60-90°F), but most do not grow well at refrigerator or freezer
temperatures. Pathogenic microorganisms may grow in foods without any noticeable change in
odor, appearance or taste. Spoilage microorganisms, including some kinds of bacteria, yeasts
and molds, can grow well at temperatures as low as 40°F. When spoilage microorganisms are
present, the food usually looks and/or smells awful.

Enzymes

Enzymes, substances naturally present in food, are responsible for the ripening process in fruits
and vegetables. Enzymes are responsible for texture, color and flavor changes. For example, as
a banana turns from green to yellow to brown, not only does the color change, but there is also
a change in the fruit'ss texture. Unblanched, frozen corn-on-the-cob may taste like the cob over
time. This is the result of enzyme action.

Air

Oxidation, a chemical process that produces undesirable changes in color, flavor and nutrient
content, results when air reacts with food components. When fats in foods become rancid,
oxidation is responsible. Discoloration of light-colored fruits can be reduced by using an
antioxidant, such as ascorbic acid or citric acid, before freezing. Vapor-proof packaging that
keeps air out helps reduce oxidation problems.

Light

Light exposure could result in color and vitamin loss. Light also may be responsible for the
oxidation of fats.

Insects, Rodents, Parasites and Other Creatures

These creatures require food to survive and damage food, making it more vulnerable to further
deterioration.

Physical Damage

Bruises and cracks on raw produce leave areas where microorganisms easily may grow.
Improperly packaged foods, dented cans and broken packages provide places for
microorganisms, air, light and creatures to enter. Gentle handling of food items will help
maintain food quality and safety longer.

Temperature

Temperature affects storage time, and food deteriorates faster at higher temperatures.
Recommended temperatures for storage areas are:

 Cupboard/Pantry 50-70°F
 Refrigerator 34-40°F
 Freezer 0°F or below

Microorganisms, both spoilage and pathogenic, grow rapidly at room temperature. To slow
microbial growth, the enzymatic and oxidation processes, store foods at lower temperatures.

Time

Microorganisms need time to grow and multiply. Other reactions, such as oxidation and
enzyme action, also require time to develop. Purchase reasonable quantities, especially of
perishable foods, to help avoid long-term storage.

CHAPTER THREE

Research methodology
3.0 INTRODUCTION

Tomato (Lycopersicon esculentum M.) belongs to the family Solanaceae is one of the most
widely grown and extensively consumed vegetable in the world. In the Nigerian Savanna, fresh
tomato is the most valuable vegetable crop. It accounts for about 18% of the average daily
consumption of vegetables in Nigeria. Tomatoes may be pearshaped, elongated, flattened and
heart shaped. They are edible, fleshy and reddish when ripe and vary in their acid composition,
with white and yellow ones being less acidic. Tomatoes can be used as savoury or flavouring in
soups and cooked foods or can be eaten as fruits. It is used in many dishes, salads, sauces and
drinks and can also be dried and ground into pancakes. Tomato fruits are mostly harvested and
transported in manners that expose them to microbial infections; they are usually harvested
and transported with locally made baskets which are prone to fungal contaminants. They are
usually displayed on benches and in baskets for prospective customers in the open market until
sold, thereby exposing them to further microbial infections beside those associated with its
surface and those from adjacent infected fruits. Fungi are the most important and prevalent
pathogens, infecting a wide range of fruits and causing destructive and economically important
losses of the fruits during storage, transportation and marketing. These study aims at
identifying fungal causing tomato rot from Gashua Main Market, Yobe State, Nigeria.

3.1 MATERIALS AND METHODS

Materials used

 Incubator
 Oven
 Petri dish
 Cotton wool
 Sterile knife
 Beaker
 Conical flask
 Sterile corks borers
  Sterile petroleum jelly
 Sterile polythene bag
 SDA plates
 SDA slants
 Weighing balance
 Germ tube
 Glass slide
 Microscope e.t.c

3.2 Samples Collection

Tomato samples were purchased from the market in Gashua. They were transported to the
chemistry laboratory in USCOEGA in a sterile polythene bags for fungal isolation. The samples
were left for one week for spoilage to occur. The spoilt tomatoes were used for the study .

3.3 Materials Sterilization

All the glass wares were first soaked and washed thoroughly with tap water and detergent
solution and then rinsed with several changes of distilled water in order to completely remove
traces of detergent and air dried completely before sterilizing them in steam oven at
temperature of 121°C for 15minutes and then allowed to cool down at room temperature
before usage. The entire working surface was also disinfected with ethanol to reduce
contamination.

All the glass wares were properly washed , dried and sterilized in the oven at 160 ° C for one
hour. The entire working surfaces were also disinfected with ethanol to reduce contaminants.

3.3.1 Samples Processing

One gram of each of the spoilt tomatoes was carefully cut with the aid of a sterile scalpel and
enriched in sterile sabouraud dextrose broth for twenty four hours . Ten fold serial dilutions of
the samples were thereafter carried out.
3.3.2 Isolation of Fungi

The pour plate method was used. One milliliter of the serially - diluted sample (103) was
dispensed into a conical flask containing sterile sabouraud dextrose agar (SDA) and two percent
chloramphenicol to inhibit bacterial growth. The contents were properly mixed and dispensed
aseptically into sterile petri - dishes. Incubation was carried out in an inverted position at 280C
for five days . The colonies that developed were counted and subcultured repeatedly on
sabouraud dextrose agar plates to obtain pure cultures. They were later stored on SDA slants
for characterization and identification .

Isolation of the fungi was carried out as described by Baiyewu et al.8 Segment (3‒5cm) of
tissues from the spoilt fruits was cut with sterile scalpel and placed on potato dextrose agar
containing streptomycin (to prevent growth of bacteria) in petri dishes and incubated at room
temperature for 5 days. Pure cultures were obtained from the isolation.

3.4 Characterization and Identification of the Isolates

The pure cultures of the moulds were identified on the basis of their colony growth pattern,
conidial morphology and pigmentation using the slide culture technique and microscopic
examination. The yeasts were characterized and identified using the Gram stain,
Chlamydospores formation, Germ tube, sugar assimilation and motility tests. The identity of
each fungus was confirmed with the aid of a mycological atlas.

3.5 Pathogenicity Test of the Isolates

The procedures of Chukwuka et al., Baiyewu et al and Onuorah et al were used. Fifteen healthy
tomatoes were properly washed with tap water, rinsed with distilled water and surface-
disinfected with ethanol. Sterile cork borers were used to bore holes in each of the tomato
fruits. Each of the isolated fungi was thereafter inoculated into the fruits after which the cores
of the fruits were replaced. Sterile petroleum jelly was used to seal the holes of the fruits to
prevent contamination. Fifteen tomato fruits wounded with the cork borers but were not
inoculated with the fungi served as controls. The inoculated tomato fruits and the controls
were placed in sterile polythene bags ( one fruit per bag). Each of the fruits was moistened with
wet balls of absorbent cotton wool to create a humid condition. The fruits were thereafter
incubated at 28 °C for five days and observed for spoilage. The fungi were re-isolated from the
fruits and compared with the original isolates. The decay rate of each fungus in the healthy
fruits was also determined by measuring its rot diameter after two weeks of its inoculation into
the healthy tomato fruit.

3 6Identification of fungi

Identification of the fungi was done according to Fawole & Oso.9 A drop of lactophenol cotton
blue stain was placed on a clean slide and with the aid of a mounted needle, a small portion of
the mycelium from the fungal cultures was removed and placed in the drop of the stain. The
mycelium was spread very well on the slide with the aid of the two mounted needles and a
cover slip was gently lowered on it. The slide was then examined under the microscope.

The observation was done at high power objective (×40) of the microscope. Morphological
characteristics of the fungi such as type of hyphae and asexual reproductive structure were
observed.

3.7 Pathogenicity of isolated fungi

Pathogenicity or decay test was carried out in order to know if the isolated fungi were really
responsible for the spoilage of tomatoes. Healthy fruits were surface sterilized with ethanol.
Cylindrical plug tissues were cut out from the fruits using a sterilized 2mm sized cork borer.
Agar plate containing a week old fungal culture were aseptically placed in these holes, then
covered and sealed off by means of petroleum jelly. The procedure was repeated separately
across each of the fungal isolates. The inoculated samples and the control were placed in sterile
polythene bags and incubated in an oven for 5 days. The point of inoculation of each type of
fungus was examined and recorded. The diameter of the rotten portion of the orange and
banana fruits was measured. The fungi were later re‒isolated from the inoculated fruits and
compared with the initial isolates.
 CHAPTER FOUR

RESULTS AND DISCUSSION OF FINDINGS

 CHAPTER FIVE

SUMMARY, DISCUSSION, CONCLUSION AND RECOMMENDATION

5.1 Discussion

The study has determined the various fungal pathogens associated with tomatoes spoilage as
well as their preliminary taxonomic grouping up to genus level. The study has also attempted to
examine the symptoms developed by these pathogenic fungi species on healthy tomato.From
the result of findings, it was seen that a total of 21 isolates were recovered from all the 25
samples of tomatoes as shown in table 2-7, constituting of six fungal genera. The study
therefore has indicated that there are a number of fungal pathogens responsible for various
diseases. According to the report of Pitt and Hocking (1985) that different genera of fungi which
are pathogenic to the fruit lives in the necrotic areas and can cause diseases in humans if
consumed directly. Fusarium species (41.4%) were the most frequently occurring fungi genera
isolated from all five samples and this is in agreement with the work of Mehrotra (2001), who
reported that Fusariumspecies were the most frequent fungal pathogenthat caused fruits’ rots
and rusts caused by fungal pathogen.
Pathogenic fungi could be attributed partly to environmental conditions prevailing in the
storage systems of vegetables. Singh (2001) had also shown that high temperature favours the
growth of these pathogenic fungi while hot and humid weather confers a most conducive
atmosphere for the growth of pathogenic fungi on vegetables. According to Singh (2001), cool
temperature and dry or low humidity reduces the rate of multiplication of fungal propagules.
This might be the reason why vegetables when kept in the refrigerator, last longer despite their
being perishable. This is true going by an earlier report of Coursey and Proctor (1975) that
metabolic activities generally decreases with decline in temperature and the activity of most
pathogenic micro–organisms diminishes with reduced temperature.

Another factor that enhances the easy penetration of fruits by these pathogens may be due to
the constant wetting to which the fruits are been subjected to by the vendors, which renders
the vegetables more susceptible to attacks. Chlorination of water used in wetting of vegetables
would reduce the extent of damage caused by the pathogens (Coplin, 1988). Also, observed,
was the mode of handling and packaging of the fruits at the market by the vendors and
consumers which is unsanitary can lead to contamination of the fruits.According to Chavan et
al. (1995), sporulation of fungal spore occurs at relative humidity above 57. 4% and
temperature of 120-300C. The water supplied to the fruit during the rainy season when the trial
was conducted was enough to aggravate the rate of sporulation and infection.

As shown in table 6, the results of the pathogenicity test that was carried out showed that all
the organisms were pathogenic and were the actual causal agents of spoilage in tomatoes. The
test also established the fact that fungi cause deterioration of fruits when they gain entrance
into them through mechanical injuries such as bruise and wounds as noted by Zitter (1985).

The isolation of these pathogens is also in line with the work ofGupta and Pathak (1985),
Baiyewu (1994), Baiyewu and Amusa (1999), Baiyewu et al. (1994, 2007) and Chukwuka et al.
(2010) who also isolatedFusarium and Rhizopus sppfrom tomatoes.

From the 50 questionnaires, 80% (40) of the respondents answered while 20% (10) of the
respondents did not answer. It was observed that most people (67.5%) use a close mode of
transportation (polyethene bags) to transport there tomatoes when purchased while a few
(22.55 and 10%) use an open mode of transportation (baskets and crates) to transport the
vegetable when purchased. Heat, microorganisms and mechanical forces are the major factors
responsible when a closed mode of transportation is used, while microorganisms, insects and
mechanical forces are factors responsible when an open mode of transportation is used. It can
therefore be said that, microorganisms and mechanical forces are the major factors responsible
for the spoilage of this vegetable as they are common to both modes of transportation. As a
result, microorganisms may gain entrancethrough physical damages caused by mechanical
forces. About 62.5% of the respondents attested to the fact that fungi are responsible for the
spoilage of tomatoes in Gwagwalada and 37.5% disagreed to this fact as shown in appendix 4.

Statistical analysis (ANOVA) had shown that fungi are greatly responsible for the spoilage of
tomatoes purchased and sold in Gwagwalada(p<0.05). This implies that there is a significant
difference.

5.2 Conclusion

This study affirmed the fact thatfungi are responsible for the spoilage of tomatoes in
Gwagwalada. Most of the categories of respondent also supported this claim from the
questionnaire conducted. From the research work, Fusarium (41.4%)species was the most
occurring pathogen while Rhizopus (4.8%)species was rarely isolated. The prevalence rate of
spoilage of tomatoes by fungi in major locations in Gwagwalada is a 100%.From the
pathogenicity test that was carried out, it was revealed thatmechanical injuries such as bruises
or cuts that occur during harvesting, post-harvesting and packing could provide infectious sites
for spoilage pathogens. These infectious sites can therefore be greatly reduced and brought to
a minimal by proper storage and handling of the vegetable.

5.4 Recommendation

From the research, to reduce the risk of afflation and other mycotoxins that are deleterious to
human health, which are produced by these fungi, it is recommended that;

The thorough washing of harvested tomatoes with clean or chlorinated water, proper cleaning
and sanitation of ware houses and disinfection of packaging containers, proper handling of the
vegetable during harvest should be done to prevent bruises and scars or other mechanical
injuries.

The inhibition of fungal growth by lowering storage temperature through storage under
refrigeration of less than 100Cbut not freezing and the use of fungicides when stored by drying.

It is important for both the farmers who harvested the tomatoes into baskets for
transportation, the marketers and consumers take necessary precaution in preventing
contamination of these tomatoes.
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