1

CHAPTER I
INTRODUCTION

Background of the Study

Fruit ripening refers to the processes that occur between the later

phases of growth and development and the time when the fruit is ready to be

consumed. Fruit ripening causes changes in the qualities of the fruit. There are

a lot of factors that cause fruit ripening. Invasion by microorganisms often

advances or hastens ripening of fruits (Brady, 2003). Ethylene is a plant

hormone widely used to ripen fruit (Digiacomo et al., 2014). The pectobacterium

Erwinia carotovora is a Gram-negative plant-specific pathogen, causing soft rot

disease of various plant hosts by degradation of the plant cell wall (Aizawa,

2014). E. carotovora increased the production of ethylene in tomatoes (Lund,

2012).

Solanum lycopersicum or commonly known as tomato is a popular fruit

vegetable extensively cultivated as a secondary crop in the Philippines, with

Northern Mindanao as one of the top major tomato-producing regions

(Department of Agriculture, 2021). However, there recently have been

complications regarding its transportation, storage, and marketing. There have

been postharvest losses of tomatoes due to rapid ripening have been occurring

at Ghana, West Africa (Camillus et al., 2023), and according to PhilStar Global,

farmers in the Cordillera Administrative Region (CAR), particularly in Ifugao,

Benguet and Nueva Vizcaya were forced to dump tomatoes amid oversupply

and lack of buyers as farmgate price reached as low as P5 per kilo. Also, a

shipment from Bukidnon, Northern Mindanao to Manila (SEARCA, 2022), and

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at Lantapan, Bukidnon (CNN, 2022) are forcing the farmers to dump their

tomatoes. This problem has been happening only recently and internationally

as well.

A current solution to this problem is wax coatings which plays an

important role in checking the storage life of fruits. It may lead to a reduction of

postharvest losses due to reduced fungal attack, transpiration, respiration, loss

of water, and flavors (Mohan et al., 2021). However, morpholine is present in

most fruit waxes and repetitive consumption of morpholine can be hazardous

to health (Muthuselvi et al, 2020). Due to this, edible coatings and films have

gained popularity for preserving fruits and vegetables (Nair et al., 2018). Edible

coatings may have the potential to inhibit the rapid quality degradation of

harvested tomatoes (Das et al., 2013). It has received more attention due to the

growing interest in reducing environmental pollution, caused by plastics, to

extend shelf life (Abebe et al., 2017). Edible coating of fruits with pure coconut

oil has been discovered to have anti-aging properties by controlling the binding

of the ethylene biosynthesis process (Nasrin et al., 2020). Extracts of both

ginger and garlic also possess antibacterial, antimicrobial and antifungal

properties that can help to extend the shelf life of fruits (Banjoko et al., 2019).

Garlic (Allium sativum L.) is a plant from the family of Aliaceae (Strika

et. al, 2017). Garlic contains at least 33 sulphur compounds like Allin, Allicin,

Trisulphide, S-allyl cysteine, Vinyl dithines Allyl propyl (Banjoko et al., 2019).

Allicin has been shown to have antimicrobial activity against many viruses,

bacteria, fungi and parasites (Strika et. al, 2017). A similar study of Fufa et al.

(2019), showed that a coating of 9.5% cornstarch mixed with 0.5% garlic bulb
 3

extract delayed the ripening of S. lycopersicum by maintaining the mature

tomato fruit for 30 days.

Zingiber officinale, commonly known as ginger, is a species in the

Zingiberaceae family. It has long been used as naturopathy due to their direct

anti-microbial activity (Islam et al., 2014). The study of Beristain-Bauza et al.

(2019) indicate that ginger contains monoterpenoids, sesquiterpenoids,

phenolic compounds, and its derivatives, aldehydes, ketones, alcohols, esters,

which provide a broad antimicrobial spectrum against different microorganisms

and make it an interesting alternative to synthetic antimicrobials. In particular,

its gingerol-related components have been reported to possess antimicrobial

and antifungal properties (Park et al., 2008).

In a similar study of Banjoko et al. (2019), bananas treated with ginger

and garlic extract delay the ripening time with a period of 16 days. In the result

obtained in their study, a higher concentration is more effective. Given that a

higher concentration has more effectivity in the ripening retardation of the fruit,

the aim of our study is to narrow down the nearest concentration that is most

effective in the higher concentration. Our study also has coconut oil as its base

to be able to create an edible coating compared to their study in which only

applied the extracts. Coconut oil also adds antioxidation effects on tomatoes.

Objectives of the Study

This study assessed the antibacterial and antimicrobial effects of ginger

and garlic extracts as ripening retardants of postharvest tomato.

Specifically, it aims to:

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1. determine the effects of Garlic (Allium sativum) and Ginger (Zingiber

officinale) extracts on the physiological and biochemical properties of

tomato in terms of:

a. weight loss

b. Percentage of fruit decay (PFD)

c. Marketable tomato fruit (MTF)

d. pH

e. Titratable acidity (TA)

2. identify the maximum number of days of the treated and untreated

tomatoes being in marketable quality;

3. evaluate the microbial activity on the surfaces of treated and untreated

tomatoes through 96-well microtiter plate-based method; and

4. assess if there is a significant difference in the quality of tomatoes in

terms of:

a. physiological properties

b. biochemical properties

c. microbial activity

Significance of the Study

This study may help reduce the postharvest losses of S.lycopersicum

due to short shelf life and invasion of microorganisms. The results of this study

will greatly help the tomato farmers and sellers in their income and will reduce

their loss. Plant extracts can act as an antimicrobial retardant of ripening and

could potentially reduce microorganism growth which is one of the causes of

ripening. The extracts are bio-efficacious and environmentally safe to use,

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which is a better alternative to chemicals. The results may also be utilized as a

basis for future research.

Scope and Delimitations

This study assessed the antibacterial ripening retardation of ginger and

garlic extracts on tomato. The fruits were hand-picked in a local farm from

Lantapan, Bukidnon by a professional classifier to get the specific maturity level

of the tomato. The data collection period was 15 days.

This study is limited to studying the antibacterial retardant effects of the

plant extracts on the ripening of tomato. It also investigates the possible

nutritional effects on the fruit itself. Included in the study are the physiological

properties in terms of; weight loss, percentage of fruit decay (PFD), and

marketable tomato fruit (MTF) and biochemical properties in terms of; pH level

and titratable acidity of the tomato. Due to lack of resources and equipment,

physiological and biochemical properties such as; color change and fruit

firmness were not investigated in the study.

The study of the experimental subject was only limited to a certain

maturity level which is the level of light red (covers 60% of the surface that

shows pinkish red or red) (Abebe, et al., 2017).

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CHAPTER 2

REVIEW OF RELATED LITERATURE

Tomato

Tomato (Solanum lycopersicum) belongs to the Solanaceae family. It

grows well over a wide range of temperature of about 18 to 30 degrees Celsius

and usually has a shelf life of 7 days at normal room temperature and

conditions. S. lycopersicum is among the most commercialized fruits due to its

high nutritional value and health-promoting compounds. (Duguma, 2021). S.

lycopersicum contains high micro and macronutrients such as vitamins,

minerals, fiber, and other beneficial compounds for human health (Mubarok et

al., 2023). Farmers are more interested in tomato production than other

vegetables due to its multiple harvests, high profitability, and potential to

increase household income and nutrition.

Even though S. lycopersicum is the source of many nutrients and health

promoting compounds, the fruit has a short postharvest life as it is a climacteric

fruit and is sold as quickly as possible (Kitinoja and Kader, 2015). Horticultural

product losses during postharvest handling in developing countries reached

50% due to storage, transportation, and packaging conditions. (Kitinoja and

Kader, 2015) Deterioration of fresh tomato fruits can result from physiological

breakdown due to ripening process, loss of water, injury, temperature or

microorganism invasion (Ahmad et al., 2020) The presence of ethylene during

tomato postharvest handling can be a major issue in preserving fruit shelf-life

by speeding up the ripening process and causing several quality changes in

fruit (Mubarok et al., 2023).

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Antibacterial for Fruit Retardation

Ethylene, a natural plant hormone, plays an important role in the ripening

of fruits. It is physiologically active in trace amounts (0.1 ppm). The capacity to

produce ethylene varies greatly among fruits. Fruits that have high ethylene

production are called climacteric. The fruits produce excess ethylene with

moderate-to-very-high production rates that lead to senescence, which causes

the fruits to have shorter shelf life (Kader, 2004). Farmers and other

researchers have been trying to address the problem of the short shelf life of

fruits which is through retardation. Low temperature storage is recommended,

but it is impractical for small-scale farmers in developing countries (Tzoumaki

et al., 2009). Another problem is that invasive bacteria grow better at higher

temperatures (Suskenik, 2012) and invasion by microorganisms often

advances or hastens ripening of fruits (Brady, 2003). One of the bacteria that

grows in tomatoes is Erwinia carotovora (Cuong, 2017) which also synthesizes

production of ethylene. This increase was directly linked to the bacterium's

secretion of pectic enzymes. These enzymes, in conjunction with those

naturally present in the plant, catalyzed the conversion of methionine into

ethylene, thereby boosting synthetic activity (Lund, 2012).

However, in recent years, there has been a surge in interest in using

edible coatings to preserve fruit quality (Tzoumaki et al., 2009). With or without

low temperature storage, edible coatings can be used to extend the postharvest

life of fresh fruits. It has the same effect as modified atmosphere storage or

packaging in that it alters the internal gas composition (Abebe, et al., 2017).
 8

Edible active coatings incorporate a wide range of functional ingredients such

as herbs, antimicrobials, antioxidants, anti-browning compounds, flavoring and

colorants compounds, and nutrients that can be used as a preservation method

to boost strategies in improving quality, safety and shelf-life of fruits and

vegetables upon storage. Such edible coatings act as physical barriers on the

fruit surface, reducing its permeability to O2, CO2, and water vapor, resulting in

lower respiration and transpiration rates and a delay in the natural physiological

ripening process (Castillo, et al., 2013). This technique has emerged as an

effective and environmentally-friendly alternative for conventional non-edible

coatings (Hassan, et al., 2020).

The addition of active agents is determined by the characteristics of the

product. Active or functional compounds include antioxidants, antimicrobials,

nutrients, vitamins, anti-browning agents, enzymes, and probiotics that can be

applied to the coating matrix to help preserve product quality (Dadpour, et al.,

2021).

Plant Extracts as Growth Retardant

In a recent study from Banjoko et al. (2019) about the influence of

aqueous extract of ginger and garlic on shelf-life studies of plantain fingers

(Musa paradisiacal) at ambient and refrigerated conditions the result obtained

showed that samples treated with 40% ginger and garlic extract are more

effective than samples treated with 20% ginger and garlic extract probably as

a result of high concentration.

In an in vitro study by Azman et al. (2020), PDA ameliorated with

aqueous ginger extract significantly inhibited 13.57% of Fusarium oxysporum,

 9

an ascomycete fungus, isolate, according to the study, evaluating the potential

aqueous ginger extract to inhibit pathogen that causes tomato wilt and delay

the weight loss of tomato fruits. With 5% and 10% concentrations of ginger

extracts, it inhibited the growth of multiple microorganisms. It also showed a

reduction of weight loss and increased firmness of tomato fruits that were

incubated for eight days at 27± 2ºC. Bacteria isolated from the tomato samples

were Bacillus, E.coli, Micrococcus, Staphylococcus, Erwinia, Pseudomonas.

Rhizopus, Penicillium, and fungi were Fusarium. Trichoderma. Aspergillus,

Cladosporium and Alternaria.

Extracts of garlic and ginger in 10% concentration were inhibitory for all

of the bacterial and fungal isolates except for Rhizopus and Aspergillus. The

10% concentration of garlic extract completely inhibited microbial growth and

was proven to be more effective than ginger extract in controlling spoilage

microorganisms, reducing weight loss, and extending shelf life. The tomatoes

were sprayed with garlic and ginger extract at concentrations of 1%, 5%, and

10%, with the highest concentration having the most effectiveness. Results

showed a reduction in spoilage, physiological loss, and microbial load on the

fruit surface (Surekha et al., 2010).

The use of natural antimicrobials has enhanced the shelf life of

tomatoes, maintaining or increasing the contents of ascorbic acid, lycopene,

and total phenolic compounds. The application of garlic and ginger extracts can

enhance the shelf life of tomatoes while being a low-cost technology that can

be utilized for fruit preservation of tomatoes. Antifungal activity of aqueous

ginger extracts against Aspergillus niger and Penicillium digitatum has been

reported (Mahajan et al., 2018). Using coconut oil as an additive with its
 10

composition film as rice starch, results showed that lipid addition to the starch

film controlled the ripening of tomatoes (Das et al., 2013).

A current solution to the rapid ripening problem is wax coatings. It plays

an important role in checking the storage life of fruits. It may lead to a reduction

of postharvest losses due to reduced fungal attack, transpiration, respiration,

loss of water, and flavors (Mohan et al., 2021). However, morpholine is present

in most fruit waxes to give thin and even films, but repetitive consumption of

morpholine as well as some other chemical agents present in fruit waxing can

be hazardous to health (Muthuselvi et al, 2020). Due to this, edible coatings

and films have gained popularity for preserving fruits and vegetables (Nair et

al., 2018). Edible coatings may have the potential to inhibit the rapid quality

degradation of harvested tomatoes (Das et al., 2013). It acts as a semi-

permeable barrier against gas and moisture, and maintains the internal quality

and appearance of food. It has received more attention due to the growing

interest in reducing environmental pollution, caused by plastics, to extend shelf

life (Abebe et al., 2017). Edible coating of fruits with pure coconut oil has been

discovered to have anti-aging properties by controlling the binding of the

ethylene biosynthesis process. It is a natural food product rich in lauric acid that

is known to add antimicrobial properties to starch-based films. Monolaurin is

one of the products of lauric acid that possesses a wide spectrum of antiviral,

antibacterial, and antifungal activities (Nasrin et al., 2020). Extracts of both

ginger and garlic also possess antibacterial, antimicrobial and antifungal

properties that can help to extend the shelf life of fruits (Banjoko et al., 2019)

Conceptual Framework
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Figure 1. Conceptual framework

Research Hypotheses

1. Null Hypothesis (H₀): There is no significant difference in the

physiological and biochemical properties of S.lycopersicum that is

treated and untreated with Garlic (Allium sativum) and Ginger (Zingiber

officinale) plant extracts.

2. Alternative Hypothesis (H₁): There is a significant difference in the

physiological and biochemical properties of S.lycopersicum that is

treated and untreated of Garlic (Allium sativum) and Ginger (Zingiber

officinale) extracts.

Operational Definition of Terms

The following terms are defined as used in the study:

Allium sativum is a plant species commonly known as garlic. Garlic has

organosulfur compounds which were proved to have antioxidant properties. Its

compounds of garlic include: alliin, allicin, ajoene , S-allylcysteine,

diallyldisulfide, diallyltrisulfide, and 1,2-vinyldithiin.

Edible coatings are thin layers or coatings applied to the surface of

fruits to preserve quality and prolong shelf life. It is like wax coating but what

differs is it is safe for human consumption. It performs the function of a semi-

permeable barrier against the movement of O2, CO2, moisture, and

 12

solutes, lowering the rate of respiration, water loss, and oxidation reaction,

which then aids in maintaining internal quality and appearance.

Ethanolic maceration is a method of extracting compounds from plant

materials by soaking them in an ethanol-based solvent. This process involves

placing the plant material in ethanol for a specific duration under controlled

conditions to extract its bioactive constituents.

Ethylene is a gas produced by some fruits and vegetables that causes

them to ripen more quickly. Some fruits are more sensitive to ethylene than

others, like climacteric fruits.

Oil-maceration is a method of extracting compounds from plant

materials by soaking them in an oil-based solvent. This process involves

placing the plant material in the oil solvent for a specific duration under

controlled conditions to allow the oil to absorb the plant’s bioactive compounds.

Plant extract is the product resulting from an extraction and separation

process where plants are used as raw material, which may help reduce the

ethylene production of some fruits.

Ripening retardation is the process of slowing down or inhibiting the

natural progression of fruit or vegetable ripening. This can be measured by

monitoring changes in color, texture, firmness, and biochemical markers

associated with ripening.

Solanum lycopersicum or commonly known as tomato is a climacteric

fruit, and due to its short shelf life, it encounters several problems in its

transportation, storage and marketing.

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Zingiber officinale is a plant species commonly known as ginger. It has

phenolic compounds which are secondary metabolites that are important

sensory and nutritional quality determinants of fruits, vegetables, and other

plants, its phenolic compounds include gingerols and shogaols.

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CHAPTER 3
METHODOLOGY

Research Design

This research was laid out in a CRD (Completely Randomized Design)

with four (4) different treatments replicated five (5) times. The tomatoes were

coated by dipping them into different concentrations: 40%, 80%, similar to the

study of Banjoko et al. (2019). Each treatment has six (6) tomato samples.

Location and Duration of the Study

The samples were observed at the residence of one of the researchers,

specifically at Flores Street, P-16, Valencia City, Bukidnon. The study was

conducted for fifteen days (15) days and was evaluated at a five-day interval.

The samples were stored in closed containers (Abd-Alla & Haggag, 2013). The

treatments were brought to the Microbiology Laboratory of The College of

Veterinary Medicine, located at Central Mindanao University for the

physiological and biochemical properties to be tested.

Figure 2. Location where the study was conducted

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Figure 3. Location of bacteria cultivation

Experimental Procedure

Figure 3. Location of bacteria cultivation

Collection and Preparation of Sample

Samples of fresh ripe postharvest tomato were collected from a local

farmer at a farm from Mabuhay and were brought to the researcher’s house

with a maturity level of light red (red covers 60% of the surface that shows

pinkish red or red), in which the maturity level of the tomatoes were determined

by a professional classifier personnel of the chosen farm. The garlic and ginger,

fresh and free from defects, were purchased in a local market and were also

brought back to the researcher's house. And coconut oil was also purchased

in a market. Each tomato fruit was washed with tap water and left air dried.

Both garlic and ginger were peeled and minced before maceration.
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Figure 4. Tomatoes collected Figure 5. Preparation of tomatoes

Oil-Maceration of Garlic

Maceration is the process of soaking plant materials in solvent. In this

study, garlic underwent oil-maceration for a faster extraction. This type of garlic

product was prepared by mixing mashed and chopped garlic into the coconut

oil (Yoshida et al., 1998). Garlic was crushed first and then remained for half

an hour exposed to air, for allicin to form and then allowed to macerate for

seven to eight days (Patil and Ravindra, 2008). After the maceration of garlic,

the macerated oil was separated from the crushed garlic by pouring it out. After

filtration, the crushed garlic was taken on a piece of cloth and all oil content

was removed by applying pressure. The concentrated part is also removed

along with the oil so it was filtered through a cloth to obtain clear oil macerate.

The oil-macerated garlic extract was placed in an airtight container, such as a

glass bottle for contamination prevention. 30mL of coconut oil was mixed per

one gram (1g) of the crude extract, making a 3.3% concentration for the stock

solution (Mansor et al., 2016).

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Figure 6. OIl-maceration of garlic Figure 7. Filtering garlic oil through cloth

Ethanolic Maceration of Ginger

Ginger underwent ethanolic maceration. Before the maceration, ginger

roots were ground to fine pieces of approximately two (2) mm size and then

extracted with 100% ethanol in the raw material/solvent ratio of 1:1, making the

ethanol 1mL/g of minced ginger root (Eberle et al., 2018). The minced ginger

roots were macerated in absolute ethanol in the beaker for 72 hours (Nguyen

et al., 2019). The ethanol extracts of ginger were filtered and filtrates were

evaporated at 40°C to dryness in a rotary evaporator. The extract obtained from

each plant material was stored in an airtight container, such as a glass bottle

at 4°C by placing it in the refrigerator to prevent contamination. 400mL of

coconut oil were mixed per ten grams (10g) of the crude extract, making a 2.5%

concentration for the stock solution (Yassen and Ibrahim, 2016).

Figure 8. Chopping of ginger Figure 9. Pouring in ethanol

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Preparation of Mixture

3300 ppm of the garlic extract was prepared from the crude extracts.

2500 ppm of the ginger extract was prepared from the crude extracts. These

were prepared by mixing the crude extract with coconut oil to make the stock

solution.

For the third treatment, a 40% concentration of the mixture was prepared

from the crude extracts. 20% of the mixture was from the crude extracts of

garlic and 20% from ginger. For the fifth treatment, a 80% concentration of the

mixture was prepared from the crude extracts. 40% of the mixture was from the

crude extracts of garlic and 40% from ginger.

Application of Extract on Tomato

The fruits were uniformly dipped into the mixture for 2-3 min. The fruits

were air dried at room temperature. When the fruits were dried, they were

placed into containers that have openings in the sides and were covered to

avoid contamination. (Abebe, et al., 2016).

Figure 10. Application of Extracts on Samples

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Bacterial Culture

Tomato rejects were collected from the same farm samples were

obtained from. Diseased tomatoes were brought to the Plant Pathology

Laboratory of the Department of Agriculture, located at Central Mindanao

University for bacterial culture. Pectobacterium (Erwinia spp.) were identified

and cultured in the tomatoes.

Figure 11. Collection of diseased tomatoes Figure 12. Bacteria cultivated

Experimental Treatment

In 500ml of coconut oil there was a 40% and 80% concentration

percentage of the oil-macerated garlic extract mixed with evaporated ethanol-

macerated ginger. The tomatoes were dipped in each of these treatments. The

table 1 shows all the mixtures of each treatment.

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Table 1. The different treatments of the study

Treatment Code Treatment Description Mixture Content

T0 (Control) Coconut Oil 500 mL Coconut Oil

T1 Pure Garlic Extracts 500 mL Garlic Extracts

T2 Pure Ginger Extracts 500 mL Ginger Extracts

T3 20% Concentration of Garlic and 20% 100 mL Garlic Extracts

Concentration of Ginger Extracts in 100 mL Ginger Extracts
Coconut Oil 300 mL Coconut Oil

T4 40% Concentration of Garlic and 40% 200 mL Garlic Extracts

Concentration of Ginger Extracts in 200 mL Ginger Extracts
Coconut Oil 100 mL Coconut Oil

Data Gathering Procedure

Data were collected through physical and disease parameters. Specific

data of parameters was recorded before treatment (day 0) and in a 5-day

interval for 15 days. Specific data to be collected are the weight, disease

incidence and shelf life.

Physiological weight loss

The fruits’ weight was weighed at day 0 treatment through storage time

and data were recorded in every 5 days interval (Tolasa, et al., 2021).

PWL(%)

where WL (%) = percentage physiological weight loss, WI=initial fruit

weight in g, and WF=final fruit weight in g at the indicated period

 21

Percentage of fruit decay

PFD was established by visually evaluating the amount of rotting and

withered tomato fruits at regular intervals and were expressed in percentages

(Tsegay, et al., 2013).

Marketable tomato fruit

MTF was assessed by counting the no. of tomatoes that were in good

condition. They were based on the firmness, appearance and quality of no

diseases present (Tsegay, et al., 2013). The MTF were expressed in

percentages of the following:

Determination of pH

The fruits were crushed, made into pulp juice, and were used to measure

the pH using a calibrated digital pH meter (Abebe, et al., 2016). The pH meter

was calibrated using pH 7 and pH 4 buffer solutions, with temperature

adjustment included in. After calibration, the electrode was rinsed twice with

distilled water, immersed in tomato juice, and the pH was measured. (Zewdie,

et al., 2022)
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Determination of Titratable Acidity (TA)

Titration was used to determine TA (expressed as a percentage of citric

acid). A 200 ml Erlenmeyer flask contained tomato juice (5ml), then water (10

ml) was added to create the fruit color light, which aided in clear end point

detection. The methyl orange indicator was then added into the flask, where it

contained 2-3 drops. Filling the 50 ml burette with 0.1 N NaOH, the 0.1 N NaOH

was carefully poured in drops into the erlenmeyer flask, and in every few drops

poured, the flask was shaken from time to time to determine if there were

changes in color. The permanent change of orange was the color of

determination that the titration had ended. The volume of NaOH solution used

for a titration was recorded and the percent titratable acidity was determined

using the following formula. (Tolasa, et al).

where: T: Titre, N: normality of NaOH, V1: volume made up, E:

milliequivalent wt. of acid, i.e. citric acid in tomato = 0.06404, V2: volume of

extract, WS: weight of sample

Determination of Shelf Life of Stored Fruits

The tomato shelf life was calculated by counting the days of the

tomatoes reaching the final stage of ripening. Collecting of the data was based

on the appearance and spoilage. (Tsegay, et al., 2013) However, the data was

only collected up to the stage where they could still remain acceptable for

commercial marketing (Tolasa, et al., 2021). A comparison was then

 23

determined between the maximum number of days of the treated and untreated

tomatoes, being in marketable condition.

Data Analysis Procedure

Before analyzing the data collected, the Shapiro-Wilk test was used to

determine if the data is normal or not. The data that was normal were analyzed

by using a parametric test called one-way analysis of variance (ANOVA) to

determine whether there is statistical evidence that the associated population

means are significantly different (Tolasa, et al., 2021). The data that was not

normal were analyzed using the Kruskal-Wallis test as a non-parametric

alternative to ANOVA. The one-way ANOVA and Kruskal-Wallis model was

used to measure the significant difference of the physiological and biochemical

effects of garlic extract and ginger extract on the treated postharvest tomatoes

with the untreated ones. A post-hoc test was no longer needed to determine

which treatment differed from the rest since it is already depicted in Figures; 13,

14, and 15.

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Chapter 4

RESULTS AND DISCUSSION

Physiological and Biochemical Properties of Tomato

The physiological and biochemical properties of the treated tomatoes

were observed and were recorded for 15 days.

a. Physiological Weight Loss (PWL)

The physiological weight loss of the tomatoes was determined for

15 days with a 5-day interval. The results of the effect of the treatments

are presented in Figure 13.

Figure 13. Results of PWL of each treatment

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The percentage of physiological weight loss increased along with the

increase of the storage duration of the tomatoes. The maximum weight loss

recorded each daily interval was 34.64% (5th day: T4), 67.44% (10th day: T0),

and 84.73% (15th day: T0). The lowest percentage was observed with T2 (5th

day: 29.41%; 10th day: 50.41%; and 15th day: 67.24).

It can be observed in Figure 13 that there was a rapid weight loss on day

5 with T4 yet it decelerated on day 10 and its weight loss became gradual until

day 15. Overall, the most rapid weight loss can be observed in T0 and T1. This

might be due to the uncontrolled maturation, acceleration of ethylene

production, and loss of moisture. This can be supported by the study of Tolasa

et al. (2021), Abebe et al. (2017), Melkamu et al. (2009) in which they state that

rapid weight loss is caused by uncontrolled ripening, sudden increase in

ethylene production and loss of moisture. In general, the fastest rate of PWL is

observed in T0 with an average of 27.22% increase per 5-day interval followed

by T1 with 25.87% increase per interval. On the other hand, the lowest rate of

PWL is observed in T2 with an average of 18.92% increase per 5-day interval.

Furthermore, the lower weight loss of fruits indicates a slow rate of

ripening and slow production of ethylene. This is also observed in the study of

Haile and Safawo (2018) and it is contrary to the findings of Tolasa et al. and

Abebe et al. This would mean that on the basis of PWL, the most effective

treatment in ripening retardation of tomato is T2 (Pure Ginger extract with

Coconut Oil) while the least effective is T1 (Pure Garlic extract with Coconut

Oil).
 26

b. Percentage of Fruit Decay (PFD)

The percentage of fruit decay of the tomatoes was determined for

15 days with a 5-day interval. The results of the effect of the treatments

are presented in Figure 14.

Figure 14. Results of PFD of each treatment

The data shown in Figure 14 indicated that the maximum PFD each daily

interval was 83.33% (fifth day: T3), and 96.67% (10th day: T2 and T3; 15th day:

T2). On the contrary, the minimum PFD perceived by T4 was recorded with the

percentages of 23.33% (5th day), 66.67% (10th day), and 86.67% (15th day)
 27

respectively. A low PFD suggests that the treatments are effective in reducing

the microbial development in the fruits, as shown by the PFD of T4.

Some treatments did not reach the 15th day interval due to the rapid fruit

decay. However, two treatments reached the 15th day interval, which are T2 and

T4. The treatments containing ginger were proven to be more effective than

other treatments which is similar to the results of Zaidi, et al (2023) but

contradicts the results of Samad, et al (2019) that had discussed that garlic was

much more effective than others. Treatments containing ginger may have been

more effective due to the fact that ethanol was used in the process of

maceration of ginger, where ethanol contains antibacterial properties, as

indicated in the discussions of Adetunde, et al (2014) and Mugao, et al (2020).

Comparing the percentage fruit decay of the two treatments (T2 and T4)

indicated a big difference between the percentages. T4 data results showed to

be the most effective in the means of PFD as the treatment induced a lower

rate of percentage fruit decay at every time interval.

Percentage of fruit decay rapidly increased during the fifth day interval

in respect to the shelf life duration of the tomatoes.This is due to the higher rate

of respiration, and higher susceptibility to infection by decay microorganisms,

compared to early harvested stages of tomato fruits, which can be supported

by the conclusions of Iqbal, et al. (2019), Bhattarai and Gautam (2006), Tsegay

et al. (2013), and Azman, et al. (2020), it pushes microorganisms to thrive,

causing deterioration and degradation through infection, eventually contributing

to a limited life span. However, the rapid increase of decay percentage is

considered to be normal as proven with the studies of Tsegay, et al. and Iqbal,
 28

et al. PFD in fruits increases and storage time of fruits decreases if the fruits

are closer to the ripened stage, compared to the fruits that are of intermediate

stage, the fruits have longer storage time.

c. Marketable Tomato Fruit (MTF)

The percentage of marketable tomato fruits was determined for

15 days with a 5-day interval. The results of the effect of the treatments

are presented in Figure 15.

Figure 15. Results of MTF of each treatment

On the initial day, tomatoes were 100% marketable. However, during

the 5th day interval, data collected showed a rapid decrease in marketable
 29

percentage. T3 had the least marketable percentage with 23.33% (fifth day

interval) and 3.33% (10th day interval), where T3 only lasted up to the 10th day

interval. The highest percentage collected was from T4 with 76.67% (5th day

interval), 33.33% (10th day interval), and 13.33% (15th day interval). The data

of T4 has shown high marketable percentages that lasted up to the 15th day

interval. A high marketable percentage indicates that it has high marketable

value and is capable of being consumed.

The MTF showed a general trend of rapid decrease during the 15 days

storage period in which similar trends were observed in the PFD and PWL,

which may have been caused by the water loss of fruits which has affected its

weight and the other parameters. The low percentage of MTF of the T2 (Pure

Ginger) and T3 (20%) was due to the developments of molds, blossom end

rots, and shriveling of fruits. These factors have caused a serious problem

during the increasing storage period of fruits as they contributed to the

reduction of MTF and PWL which has been experienced by all the fruits in

treatments. The deterioration of fruits caused by the factors was also

experienced by Tigist, et al (2012) and Munir and Iqbal (2023). However, in the

case of T2 (pure ginger treatment), shriveling was mostly present throughout

the day interval and had less presence of microbial invasion. Throughout the

storage period, although the ginger had 3.33% on the 15th day interval, it was

visually evaluated that the pure ginger treatment had the best appearance out

of all treatments. The reason why it had a decrease of MTF was because most

of the fruits were shrivelled which contrasts to the decrease of MTF of other

treatments because of microbial invasion. According to the results of Azman,

ginger extracts were able to reduce the fungal spoilage of the tomato fruits
 30

which supports how there was less microbial invasion in the fruits with ginger

extracts.

The high percentage of MTF coated with T4 (40% garlic and ginger) on

the 5th day interval significantly connected with its capacity to limit the

percentage of decay of tomato fruits by acting as a physical barrier for the entry

of microbes and its potential in reducing weight loss the same as discussion of

Sharmin et al (2015) Therefore, the postharvest treatments have reduced

weight loss, excessive shrinkage, and spoiling typically improve the fruit

marketability.

d. pH Level and Titratable Acidity (TA)

The pH level and titratable acidity of the tomatoes was

determined during the 15th day of observation. The results of the effect

of the treatments are presented in Table 2.

Table 2. Average pH level and TA of each treatment

The pH level and TA of the tomatoes were collected on the 15th day of

observation in which the tomatoes are considered fully ripe. It is observed in

the data that as pH level of the tomato goes higher, its TA goes lower. The data

shows that the pH level of the tomatoes ranges from 3.9 - 5.32. Highest pH

level can be observed in T3: 5.32 (40% extracts) followed by T1: 5.34 (garlic

extract). These two treatments are the only ones who exceeded pH level 5. The
 31

lowest pH can be observed in T2: 3.90 (ginger extract) followed by T4: 4.05

(80% extracts) and T0: 4.53 (coconut oil).

While the titration acidity expressed as a percentage of citric acid (TA)

ranges from 0.15% - 0.85% with T3 (0.15%), T1 (0.17%), T0 (0.25%), T2

(0.65%), and T4 (0.85%) in increasing order. It can be observed that despite

not having a trend, the tomatoes with higher pH have a lower TA. Therefore,

the pH level of the tomatoes is inversely proportional to its TA. This observation

is similar to the study of Abebe et al. (2017), and Bhattarai and Gautam (2006).

At harvest, the pH of the tomatoes that were collected resulted in an

average of 4.2. The pH of fruits increases as fruits undergo ripening. This can

also be observed in the study of Banjoko et al. (2019), Melkamu et al. (2009),

and Tolasa et al. (2021). Based on our results, it is applicable in treatments T0,

T1, and T3. However, it can be observed that there is a decrease in pH in

treatments T2 and T4 which was also observed by the study of Olaniran et al.

(2020). The study suggests that low pH could enhance the stability of bioactive

compounds during storage, thus extending shelf life. This can be verified by

the physiological results in our study where mostly T2 and T4 have better

results than the rest.

Shelf Life

The shelf life determines the quality and marketability of the fruits. It is

critical for the determination of its maximum storage and the marketing of fruits.

The maximum storage life of the treated and untreated tomatoes being in

marketable quality was identified within 15 days. As reflected by the presented

physiological parameters, it can be determined that the treatments that has

 32

reached the maximum shelf life were T2 (pure ginger) and T4 (40% ginger and

garlic extracts) which reached 15 days, followed by the T0 (pure coconut oil),

T1 (pure garlic extracts), and T3 (20% garlic and ginger extracts) which only

reached up to 10 days respectively. This proves that T2 and T4 treatments are

effective in extending the shelf life of the tomatoes. As discussed in the PFD

and MTF parameters, the extracts containing ginger were more effective in

reducing the microbial activity, moreover the fact that there was ethanol present

in the ginger extracts contributed to its shelf life as ethanol contains

antibacterial properties, whereas these findings can be supported by the results

obtained by Azman, et al (2020), Adetunde, et al (2014) and Mugao, et al

(2020). With the treatments exceeding the normal shelf life of tomatoes of 7

days, we can therefore conclude that treatments were effective in extending

the shelf life.

Antibacterial activity

The antibacterial activity of each treatment was tested using

Pectobacterium carotovora as its bacteria.

Table 3. Results of antibacterial activity of each treatment

The results show that the zone of inhibition of all the treatments is zero

(0) mm. This might be caused by the coconut oil being the solvent of all the

treatments. Microorganisms, especially pectobacterium, are highly susceptible

to coconut oil. This can be confirmed by the study of Abbas et al. (2017) and a
 33

more recent study of Balea et al. (2023).

Significant difference in the quality of tomatoes

a. physiological properties

Physiological weight loss (PWL)

Using the Shapiro-Wilk test, data collected on all days are normal

therefore analysis of variance, or ANOVA was used to determine the

significant difference. ANOVA revealed that the different pre-storage

treatments had a highly significant effect on the percentage of

physiological loss on tomato fruit weight. It showed that in every 5-day

interval, there was a significant difference between the treatments.

Percentage of fruit decay (PFD)

Using the Shapiro-Wilk test, data collected on the 5th and 10th

day are normal and on the 15th day is not. ANOVA showed that the data

on the 5th and 10th day has a significant difference between the

treatments. Since the data in the 15th day was not normal, Kruskal-

Wallis was used to determine the significant difference which revealed

that there was no longer a significant difference between the treatments.

Marketable tomato fruit (MTF)

Using the Shapiro-Wilk test, data collected on the 5th and 10th

day are normal and on the 15th day is not. ANOVA revealed that there

was a significant difference between the treatments in the data on the

5th day only. On the 10th day, the p-value resulted in 0.237 which

indicates that there is no significant difference between the treatments.

 34

Since the data in the 15th day was not normal, Kruskal-Wallis was used

to determine the significant difference which revealed that there is no

significant difference between the treatments.

b. biochemical properties

pH level and titratable acidity (TA)

Since the data only have 5 observations, the Kruskal-Wallis test

was used to test the significant difference between the pH level and TA

of the pulp of tomatoes. Both of its p-values resulted in 0.406. This

indicates that there is no significant difference between the pH level and

TA of the tomatoes.

c. microbial activity

A statistical analysis was not conducted in this parameter since

all the treatments had the same results.

 35

Chapter 5

SUMMARY, CONCLUSION, RECOMMENDATION

Summary

The results showed that the maximum weight loss recorded was

observed in T0 while the minimum was observed in T2. The data has shown that

the maximum PFD each daily interval was observed in T2. On the contrary, the

minimum PFD was recorded in T4. T3 had the least marketable percentage

and it only lasted up to the 10th day. The data of T4 has shown the highest

marketable percentages that lasted up to the 15th day interval. The data shows

that the pH level of the tomatoes ranges from 3.9 - 5.32. Highest pH level can

be observed in T3 while the lowest pH can be observed in T2. The shelf life of

our results demonstrate that the treated tomatoes reached their maximum

marketable quality within 15 days. Treatments T2 (pure ginger) and T4 (40%

ginger and garlic extracts) exhibited an extended shelf life of 15 days. For the

antibacterial activity, all treatments showed no zone of inhibition against

pectobacterium.

Conclusion

This study revealed the antibacterial and antimicrobial effects of ginger

and garlic extracts as ripening retardants of postharvest tomato.

The physiological weight loss (PWL) increased with storage duration,

pure ginger treatment with the least amount of weight loss indicating slowest

rate of ripening retardation among treatments. Treatments containing ginger

extract showed slower weight loss, suggesting better ripening retardation.

Similarly, the percentage of fruit decay (PFD) was lower in treatments

 36

containing ginger, due to its antibacterial properties. Marketable tomato fruit

(MTF) percentages declined over time, with ginger extract-treated fruits

exhibiting better appearance and reduced microbial invasion. The pH level and

titratable acidity (TA) revealed varying ripening stages and potential shelf-life

extension in treatments with lower pH.

Our results demonstrate that the treated tomatoes reached their

maximum marketable quality within 15 days which suggests the effectiveness

of these treatments in prolonging tomato shelf life.

Assessment of physiological parameters, including physiological weight

loss (PWL), percentage of fruit decay (PFD), and marketable tomato fruit

(MTF), revealed significant differences among treatments. Evaluation of

biochemical properties such as pH level and titratable acidity (TA) did not reveal

significant differences among treatments. This indicates that treatments did not

significantly alter the biochemical composition of tomato pulp. Regarding

microbial activity, the antibacterial activity of treatments against Pectobacterium

carotovora was assessed. All treatments exhibited zero inhibition zones,

attributed to coconut oil, a common solvent in the treatments, due to its high

susceptibility against pectobacterium.

Our findings highlight the efficacy of the treatments containing ginger

extract in maintaining tomato quality and extending shelf life of tomato.

Recommendation

Promotion of these edible coatings needs to be increased among the

consumers so as to broaden and diversify their use in fruits and vegetable

 37

preservation. Based on the results of the study, it is recommended for future

researchers interested in this field to conduct this study with a larger sample

size and more concentrations of the extracts. Also, it is recommended for future

researchers to evaluate more physiological and biochemical properties of the

chosen fruit for further indications and reasons of the ripening retardation

effects of the extracts. It is also recommended to include an antioxidant capacity

test on the treatments as ginger and coconut oil are high in antioxidants.