COLLEGE OF SCIENCE AND TECHNOLOGY

SCHOOL OF SCIENCES
DEPARTMENT OF BIOLOGY
OPTION: BIOCHEMISTRY
ACADEMIC YEAR: 2021-2022

Final Research Project

THE ANTIMICROBIAL AND PRESERVATIVE ACTIVITY OF ALLIUM SATIVUM

(GARLIC) ON SOLANUM LYCOPERSCUM (TOMATOES)

Final project dissertation submitted in partial

fulfilment of the requirements for a
Bachelor’s of Science degree in Biology,
option of Biochemistry

By: - Seth NIYONKURU

Reg. number: 220004000
- Fabrice ABIKUNDA
Reg. number: 220002312
- Brian KAMUGISHA
Reg. number: 220013623

Supervisor: Dr. Diane UMUHOZA

Kigali, April 2023

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I. DECLARATION
We, Fabrice ABIKUNDA, Brian KAMUGISHA and Seth NIYONKURU hereby declare that
this research project for the award of a Bachelor of Science Degree in Biology - Biochemistry,
with the title "The Antimicrobial and Preservative Activity of Allium sativum (garlic) on
Solanum lycopersicum (tomatoes)” in Kigali city, is the outcome of our original preparation
and to the best of our knowledge. Except when appropriate acknowledgment has been made in
the text, no part has been published or presented for a different degree at this university or
elsewhere.

Fabrice ABIKUNDA (REG.NO: 220002312)

Signature: 2nd May , 2023

Seth NIYONKURU (REG.NO: 220004000)

Signature: 2nd May, 2023

Brian KAMUGISHA (REG.NO: 220013623)

Signature: 2nd May, 2023

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II.APPROVAL
This is to approve that the final year project entitled “The Antimicrobial and Preservative
Activity of Allium sativum (garlic) on Solanum lycopersicum (tomatoes)” is the work of Seth
NIYONKURU, Fabrice ABIKUNDA and Brian KAMUGISHA, the students at the University of
Rwanda, college of Science and Technology (UR-CST). The information provided is based on
different studies from worldwide researchers and direct observations during laboratory
experiments.

Supervisor : Dr. Diane UMUHOZA

Signature………………………………..

Date: 2nd May 2023

Head of department : Dr Diedonne MUTANGANA

Signature……………………………………………….

Date : 2nd May 2023

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III. DEDICATION
We are willing to dedicate our project to

 Almighty God
 Our parents
 Our supervisor: Dr. Diane UMUHOZA
 Lecturers of School of science especially in Biology department
 Our classmates
 Our friends
 Any other person who gave us his/her support of any kind.
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IV. ACKNOWLEDGEMENT

We first give thanks to the Almighty God for his mercy and safeguarding.

Second, we would like to express our gratitude to our supervisor, Dr. Diane UMUHOZA, for her
patience, support, and readiness to help us complete this research project.

We appreciate the workers at various institutions for letting us use their facilities and the
members of the community who provided us with important information.

Our thankful gratitude is made to UR-employees CST's and the department of Biology, for their
advice and all facilitations.

We appreciate the assistance of laboratory technicians.

Finally, we would want to express our sincere gratitude to all of our classmates, families, and
close friends for their encouragement and support during this time.
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Contents
I. DECLARATION................................................................................................................................i

II.APPROVAL...........................................................................................................................................ii

III. DEDICATION....................................................................................................................................iii

IV. ACKNOWLEDGEMENT.................................................................................................................iv

V. LIST OF FIGURES............................................................................................................................vii

VI. LIST OF TABLES............................................................................................................................viii

VII. LIST OF ABBREVIATIONS AND ACRONYMS.........................................................................ix

VIII. ABSTRACT......................................................................................................................................x

CHAPTER 1. GENERAL INTRODUCTION........................................................................................1

1.1 Background and Literature review....................................................................................................1

1.1.1. Types of Allium sativum (garlic) and their differences....................................................1

1.1.2. Uses of Allium sativum.......................................................................................................2

1.1.3. Mechanisms of action of garlic extract as antimicrobial agent.......................................3

1.1.4. Introduction to tomatoes and their rotting mechanism..................................................4

1.3. Objectives...................................................................................................................................9

1.3.1. General objectives..............................................................................................................9

1.3.2. Specific objectives..............................................................................................................9

1.4. Research hypotheses..................................................................................................................9

1.5. Research questions.....................................................................................................................9

1.6. Significance of study................................................................................................................10

CHAPTER 2. METHODOLOGY..........................................................................................................11

2.1. Study area description..................................................................................................................11

2.3. Preparation of aqueous garlic extract.........................................................................................11

2.4. Testing antimicrobial activity of Allium sativum........................................................................14

2.5. Preservative activity of Allium sativum against spoilage of tomatoes.......................................15

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2.6. Data analysis.................................................................................................................................15

CHAPTER 3. RESULTS........................................................................................................................16

3.1. Preparation of aqueous garlic extract.........................................................................................16

3.2. In vitro antimicrobial properties..................................................................................................17

3.3. Preservative activity of Allium sativum on fresh tomatoes.........................................................19

CHAPTER 4. DISCUSSIONS................................................................................................................24

CHAPTER 5. CONCLUSION AND RECOMMENDATIONS...........................................................29

5.1: Conclusion.....................................................................................................................................29

5.2: Recommendations........................................................................................................................29

REFERENCES........................................................................................................................................30

APPENDIX................................................................................................................................................A
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V. LIST OF FIGURES
Figure 1: Multiple functions of garlic .............................................................................................6

Figure2: Images indicate (A) WGS bulbs, (B) PGS bulbs, (C) WGS cloves and (D) PGS cloves
.......................................................................................................................................................13

Figure 3: Preparation of garlic aqueous solution...........................................................................16

Figure 4. Images that identify the growth inhibition zone due to antimicrobial activity of both
PGS & WGS with their respective positive and negative control.................................................17

Figure 5. Graph represent the length of growth inhibition zone....................................................18

Figure 6: the 3-days’ time interval of how half-cut tomatoes spoiled over time. The number in
brackets stands for days of preservation. PGS & WGS stands for purple garlic species and white
garlic species respectively.............................................................................................................19

Figure 7: the image that show day 9 for half-cut tomatoes under investigation............................20

Figure 8: Image of coated and uncoated tomatoes for first 6 days and last 9 days. Note: that the
number in bracket stands for the preservation day........................................................................20

Figure 9: The percentage weight loss of each sample...................................................................22

Figure 10: The graph indicating how mass decreased over time during preservation.................22

Figure 11. The chemical structure of alliin, allicin and its derived OSCS.....................................A

Figure 12. Image showing global postharvest loss of tomatoes.....................................................C

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VI. LIST OF TABLES

Table 1: Hierarchical Taxonomy of garlic ......................................................................................6

Table 2. Materials and reagents used in extraction and preparation of coating.............................12

Table 3: Variation of growth inhibition provided by antimicrobial agents...................................18

Table 4: Decrease in a mass of preserved tomatoes over time and their corresponding standard
deviation........................................................................................................................................21
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VII. LIST OF ABBREVIATIONS AND ACRONYMS

1. AGE: Aged Garlic Extract
2. MPFVs: Minimally Processed Fruits and Vegetables
3. SAC: S-Allyl Cysteine
4. GE: Garlic Extract
5. PGS: Purple Garlic Species
6. WGS: White Garlic Species
7. SAMC: S-Allylmercapto-L-Cysteine
8. EPS: Extracellular Polymeric Substances
9. OSCS: organosulfur compounds
10. QS: quorum sensing
11. GAP: Good Agricultural Practices
12. cm: centimeter
13. PG: Polygalacturonase
14. PEL: Pectate Lyase
15. PME: Pectin Methylesterase
16. CSAM: Chain Analysis and Commodity Systems Assessment Methodology
17. LAB: Lactic Acid Bacteria
18. GEO: Garlic Essential Oil
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VIII. ABSTRACT
Many years ago, garlic (Allium sativum) has been used as culinary spice and therapeutic agent by
our ancestors. In the second world war, soldiers used it as antimicrobial agent which helped them
to heal wounds. Since 1990 scientific researchers started studying how garlic can be processed in
order to be used in different activities such as therapeutics, culinary activities, meat
preservatives, preparation of edible coatings and films for preservation of vegetables and fruits.
In this study, we focused on the Antimicrobial and preservative activity on fresh tomatoes using
two varieties Purple Garlic Species (PGS) from hardneck garlic type and White Garlic Species)
from softneck garlic type. antimicrobial activity was tested by inoculating general
microorganisms on nutrient agar plates and record growth inhibition zone in comparison of
garlic extract and chemical antibiotics. Coatings were prepared by mixing starch with garlic
extract Whole tomatoes and half-cut ones were dipped in coatings for 10 minutes and then stored
at room temperature. It was observed that the coatings prolonged the shelf life of tomatoes
compared to their corresponding control. Another observation is that PGS showed more
effectiveness than WGS coatings.
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CHAPTER 1. GENERAL INTRODUCTION

1.1 Background and Literature review

Among all domesticated plants, Allium sativum (garlic) is one of the oldest and has been used
medicinally for thousands of years. Allium is a genus name that derives from the Celtic word
"all," which means pungent. The term "garlic" has several possible origins. Gar, which means
"spear" (because the shape of a clove resembles the head of a spear), and leac, which means
"leek," are Anglo-Saxon terms for the spear or gar-leac plant. It appears that garlic originated in
central Asia before migrating to China and other nations around the world (Saif et al., 2019).

1.1.1. Types of Allium sativum (garlic) and their differences

There are more than 600 cultivars of garlic cultivated, according to some sources. The majority
of them fall into one of two broad categories: Sativum, or soft-necked garlic, and Ophiosco-
rodon, or hard-necked garlic (also known as "ophio" garlic). Hard stemmed flower stalks bearing
aerial bulbils are produced by hard-necked garlic varieties like Purple Stripe and Porcelain. A
hard-necked cultivar called rocambole has a distinctively curled bloom stalk. Typically, soft-
necked garlics like Artichoke and Silverskin don't grow flower stalks. Each type of garlic has
unique growing conditions and, to the discerning palette, a distinct flavor (Wayne J. McLaurin,
Adams, David, 2015). It is important to note that softneck garlic are usually white whereas
hardneck garlic can be brown, red, purple and white in color.

(Miller, 2021) have provided some of the differences between hardneck and softneck garlic
which include: Large, oval to triangular-shaped cloves, usually 4 to 12, are produced in a single
layer by hardneck garlic plants while softneck heads are often larger and contain 8 to 20 cloves
on average, many of which are shaped erratically. The majority of hardneck garlic types have
easily removable skins while the uneven shape and tight, thin skin of softneck cloves make
peeling more challenging. Moreover, this has an impact on shelf life, with softneck kinds storing
much better. While softneck variants do better in regions with warm winters, hard neck varieties
are more resilient in colder locations.
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1.1.2. Uses of Allium sativum

Allium sativum is an amazing plant with numerous health benefits, including antibacterial,
antithrombotic, hypolipidemic, anti-arthritic, and anti-tumor activity (Thomson & Ali, 2005).
Apart from its therapeutic properties, around 65% of a garlic bulb's nutritional makeup is made
up of water, followed by 28% carbohydrates, 2.3% organosulfur compounds, 2% protein, 1.2%
free amino acids, and 1.5% fiber. 100 grams of garlic contain approximately 149
kilocalories when consumed (Suleria et al., 2015). Many chemical components are included in
Allium sativum, including 17 amino acids, more than 33 Organosulfur Compounds (OSCs), 8
minerals (calcium, potassium, magnesium, germanium, copper, zinc, and iron), vitamins (A, B1,
B2, B3, B6, B12, C, D, and E), and a few enzymes, including allinase and peroxidase (Foroutan-
Rad et al., 2017). This makes it a priceless culinary spice that is used as a seasoning for
vegetables, sauces, and a variety of meat preparations. Garlic has been used as a flavoring and
food ingredient in cooking all over the world because it improves the flavor and aroma of a wide
variety of foods (Saif et al., 2019).

Not only that, because the experiment's findings demonstrated that garlic might be utilized as an
efficient natural meat preservative to stop meat from spoiling as a result of bacterial growth
(Sarma, 2004). According to (Benkeblia, 2004), substantial research has been done on the
antibacterial and antifungal activity of garlic against a variety of Gram-positive and Gram-
negative bacteria. Also, he stated that 15 IU of penicillin is equivalent to 1 mg of allicin, which is
a (+)-S-methyl-l-cysteine sulfoxide, in terms of antibiotic activity. (Phan et al., 2019) also stated
that the antibacterial activity results are consistent with those of the other researchers who found
that aqueous extracts of fresh garlic cloves or commercially available freeze-dried garlic powder
had an inhibitory effect on food-related bacteria, yeasts, fungi, and viruses. The ethanolic
extracts of clove and garlic in the study made by (Kumar Pundir et al., 2010) demonstrated
inhibitory efficacy against all six investigated food-associated bacteria, with the diameter of the
growth inhibition zone varying between 25 and 32 mm (in clove) and 20 and 31 mm (in garlic).
In this study, we shall focus on antimicrobial and preservative properties of Allium sativum
(garlic). garlic solution is used to prepare edible coatings which prolong the shelf life of fruits
and vegetables to maintain and protect the quality of postharvest (Desta Dugassa, 2021). Allicin
(diallyl thiosulfinate), which makes up the majority (60 to 80%) of the bioactive chemicals in
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garlic is the main active ingredient responsible for garlic's antibacterial abilities and thus the
preservative activity (Pranoto et al., 2005).

1.1.3. Mechanisms of action of garlic extract as antimicrobial agent

Allicin's primary antimicrobial impact is brought on by an oxidative interaction with vital thiol-
containing enzymes (Jabar & Al-Mossawi, 2007). Bacterial biofilms are collections of bacterial
cells embedded in a matrix of secreted lipids, proteins, and nucleic acids known as extracellular
polymeric substances (EPS). Quorum sensing (QS) signaling is a complicated process that is
involved in the production of biofilm. The utilization of numerous phytochemicals, such as garlic
and its organosulfur compounds, as well as synthetic antibiofilm agents has drawn a lot of
attention. Garlic and its components have been used in antibiofilm and anti-QS investigations.
Using in vitro experiments, it was discovered that Aged Garlic Extract (AGE) inhibited S.
aureus's coagulase activity (Bhatwalkar et al., 2021).

The fact that different sulfhydryl enzymes can be inhibited by allicin suggests that allicin's
antibacterial effect involves reacting with the sulfhydryl groups of several metabolically
significant bacterial enzymes. This helps sulfur compounds from garlic to bind to the enzymes
that are used in cell wall and cell membrane synthesis and prevent them from performing their
activities. This then, results in failure in synthesizing cell wall which finally cause an osmotic
imbalance in a bacterial cell and it dies. Strong food pathogen inhibitors include garlic. Foods
that are pathogen-contaminated could be harmful to consumer health. Garlic edible coating use
would lengthen the shelf life and reduce the risk of spoilage and food poisoning in processed
foods (Kumar & Berwal, 1998). Although DNA and protein synthesis are also somewhat
hindered, RNA synthesis is the principal mechanism by which allicin demonstrates its
antimicrobial activity. This suggests that RNA is the main target of allicin action (Yadav et al.,
2015).

Despite the large number of commercially available garlic supplements, they can be divided into
four groups according to the mode of preparation: dehydrated garlic powder, garlic essential oil,
garlic oil macerate, and raw garlic extract (also known as aged garlic extract (AGE) with
extended time) (Milner, 2001). The concentration of allicin in garlic juice extract is highest,
followed by garlic powder and fresh garlic. The processing of AGE is distinct from that of the
other three varieties of garlic products. This extract is matured for up to 20 months, as the name
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suggests. During this aging process, the odorous, harsh and irritating chemicals in garlic are
transformed naturally into stable and harmless sulfur compounds. In addition to a number of oil-
soluble sulfur compounds, aged garlic extract largely contains water-soluble sulfur compounds
such S-Allyl Cysteine (SAC) and S-Allylmercapto-L-Cysteine (SAMC). Because SAC is
bioavailable and its concentration rises with extraction and age, it can be utilized for
standardization. Also, numerous toxicological investigations have supported AGE's safety
(Milner, 2001).

Garlic is a preservative because of its antibacterial properties. Literature shows that treatments
containing 5, 10, and 15% by weight (by wt.) of fresh garlic segments, ground with fresh lean
camel meat, increased the period of shelf-life by two, three, and more than four times,
respectively, compared with the corresponding control samples, regardless of storage
temperature whether fridge temperature is between 2–3 °C, incubator under 12°C or at room
temperature 20–22 °C (Al‐Delaimy & Barakat, 1971). Studies show that garlic and ginger
worked well together as bio-preservatives to lower bacteria and yeast counts. Ginger at the same
dose was less efficient than garlic at 2% and 4% against Lactic Acid Bacteria (LAB) and yeast
load. 4% of garlic was added to tomato paste to reduce chemical features, prevent
microbiological deterioration, and make it more palatable to the taster (Olaniran et al., 2015).

1.1.4. Introduction to tomatoes and their rotting mechanism

The tomato, (Solanum lycopersicum L.), is the third most frequently planted vegetable crop in
the world. Worldwide, household consumption is rapidly rising. Tomato is of juicy flesh
endocarp belonging to the fruit class, berry. It naturally contains high levels of protein, dietary
fiber, vitamins, and minerals. In addition to serving as food, tomatoes are also used as a
medication, dietary supplement, flavoring agent, and human system cleanser (Omolaran et al.,
2016). (Rosales et al., 2009) stated that plant cell walls contain a lot of pectins, which make up
around one-third of the overall composition. Due to their excellent capacity as a gelling agent,
these fibers have a significant influence on the consistency of fruits as well as in the textural
changes during ripening, storage, cooking, or other procedures. Consequently, the enzymatic
breakdown of tissues and the solubilization of protopectins are principally responsible for the
tissues' softening. Many enzymes that hydrolyze the component polymers cause the cell wall to
deteriorate in most cases. The pectolytic enzymes polygalacturonase (PG), pectin methylesterase
 5

(PME), and pectate lyase (PEL) have been the subject of the most research in tomato fruit, which
causes significant losses in the texture and viscosity of the fruits throughout their processing,
leading to a decrease in market quality. A deficiency of Ca 2+ in fruit also can lead to blossom end
rot (BER) because it was indicated that all tomatoes attacked by BER had a low quantity of Ca 2+
in their content (Rosales et al., 2009).

Apart from those internal factors that contribute to tomato rotting, there are other external factors
including microorganisms which can accelerate tomato spoilage. (Etebu et al., 2013) reported
that bacteria and fungi are the two main groups of microorganisms that contribute to tomato
deterioration. Tomatoes are not gradually deteriorated by other plant pathogens, such as viruses
and worms, but they may cause infections and losses after harvest. (Omolaran et al., 2016) stated
that both bacteria (i.e. Staphylococcus aureus, Bacillus spp,Bacillus subtilis, Bacillus cereus,
Bacillus Aureu, Lactobacillus fermenti, Escherichia coli, Pseudomonas stutzeri, Leuconostoc spp
and Rothia spp, Klebsiella aerogenes, Salmonella typhi, Pseudomonas aeruginosa) and fungi
(i.e. Aspergillus flavus, Rhizopus stolonifer, Aspergillus fumigatu, Penicillium expansum,
Penicillium notatum, Mucor mucedo, Monilia spp, Proteus mirabilis, Rhizopus stolonifer,
Botrytis cineria, Saccharomyces spp, Rhodotorula spp and Verticillium albo-atrum) are
responsible for tomato fruits spoilage by contaminating them during field cultivation, harvesting,
post-harvest handling and distribution which finally degrade tomatoes protective cover
(epidermal layer) rendering to spoilage.
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Table 1: Hierarchical Taxonomy of garlic (Khorshed Alam et al., 2016)

Domain Eukaryota
Kingdom Plantae
Subkingdom Tracheobionta
Super division Spermatophyta
Division Magnoliophyta
Class Equisetopsida
Subclass Magnoliidae
Superorder Lilianae
Order Asparagales
Family Amaryllidaceae
Genus Allium
Species Sativum (Allium sativum)

Figure 1: Multiple functions of garlic (Foroutan-Rad et al., 2017)

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1.2. Problem statement

Due to their nutritional value and positive impacts on human health, fruits, vegetables, and
cereals are essential components of the human diet. Good sources of vitamins, minerals,
phytochemicals, dietary fiber, and antioxidants can be found in fruits and vegetables (Alegbeleye
et al., 2022,)(Zhao et al., 2020). Since their increased use in prepared mixed salads as a source of
fresh, healthful, and convenient food, consumers' perceptions about minimally processed, fresh
fruits and vegetables (MPFVs) have changed significantly (De Corato, 2020). However, those
fruits face the problem of being spoiled by microorganisms from different areas. Microorganisms
that cause spoilage can infect the crop when it is planted, as it grows in the field, when it is
harvested and handled afterward, or when it is stored and distributed (Barth et al., 2009).
Additionally, it is predicted that roughly 20% of all cultivated fruits and vegetables are spoiled
each year (Barth et al., 2009).

Post-harvest loss worldwide: Kitinoja et al., (2018) in their article titled “A review of global
postharvest loss assessments in plant-based food crops: Recent findings and measurement gaps”
showed that with 350 to 400 data points collected for each crop, postharvest loss of fruits and
vegetables in Bangladesh ranged from 23.6 to 43.5%. Jackfruit (43.5%) had the highest overall
loss rate, followed by pineapple (43.0%), papaya (39.9%), and cauliflower (34.4%). Red
amaranth (28.8%), cucumber (27.1%), mango (27.4%), eggplant (29.4%), okra (32.3%), and
tomato (32.9%) all had mid-range losses. Orange (23.6%), banana (24.6%), and litchi (24.9%)
postharvest losses were less than average. They also identified that in Trinidad and Tobago and
Guyana, where cassava was deemed "unfit for sale," an FAO research from 2015 evaluated the
total estimated economic losses in cassava at US $500,000 and US $839,000, respectively.
According to one study, which called for the market prices to be monitored throughout time,
there were financial losses for tomatoes and cauliflower per dealer, wholesaler, and retailer. For
vegetable crops in Cambodia, Gautam and Buntong (2015) reported on economic losses per
hectare. For tomatoes, the economic loss was projected to be at $4,213 USD per hectare and
$2,208 USD for green mustard.

(Branthôm, 2023) stated that the FAO reported that global tomato production for both processing
and fresh consumption reached just over 189.1 million metric tonnes in 2021, up 2% from the
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184.8 million metric tons (MMT) grown in 2020 and 4% from the average (182.7 MMT) of the
previous three years (2018-2020). According to (RAB et al., 2018), One of the most widely
consumed vegetables in Rwanda is the tomato, which is offered both fresh and processed on the
domestic market. Tomato processors are expanding domestic demand. Production has increased
from 135,000 tonnes in 2010 to 154,000 tonnes in 2014. Mechanical damage grew from the farm
(33%) to the retail market (68%) linearly. Overall, 21% of tomatoes were thrown away at the
farm level, 11.5% at the collection point, 10% at the wholesale market, and 13.6% at the retail
market, resulting in a total of 56% of tomatoes being thrown away in their travels from farm to
market.

For instance, the Chain Analysis and Commodity Systems Assessment Methodology (CSAM)
team found significant damage (between 20 and 60%) during harvest at the farm. The market
value of the tomato crop dropped as the quality rose, creating a sizable window for post-harvest
handling improvements and container investments. Tomatoes of excellent quality could be sold
for between 1000 and 800 Rwf per kg. In Nyabugogo, Rwanda (March 2023) as an illustration:
Soft fruits are worth 800 Rwf per kg, but damaged or broken fruits are worth 500 Rwf per kg. If
the yearly tomato production is 154,000 tonnes, the market value will decline from
USD111,493,212.67 to USD 69,683,257.9 million (which is 62.49% percentage loss) annually.

For this reason, Good agricultural practices (GAP) should be used as early intervention methods
during crop development and harvesting to significantly reduce production loss due to spoiling at
all stages of the food-to-fork continuum (Barth et al., 2009). Apart from decrease in productivity
and expensive preservatives, spoiled fruits and vegetables are likely to cause diseases. Typhoid,
paratyphoid, and salmonellosis can all be brought on by salmonella. Clostridium botulinum is the
most frequent cause of botulism. Shigella, Yersinia, Cryptosporidium, Cyclospora, Hepatitis A
virus, and other pathogens can also result in foodborne sickness.

The use of edible protective films and coatings in packaging is one approach for prolonging the
shelf-life of lightly perishable fresh and MPFVs. Since the 1990s, research on edible films and
coatings that can prevent the growth of pathogenic bacteria in food has been thought to be a
particularly promising area of study for new natural packaging films comprised of biodegradable
and edible materials (De Corato, 2020).
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1.3. Objectives
1.3.1. General objectives
The main objective of this study is to determine the antimicrobial and preservative activity of
Allium sativum (garlic) on fresh tomatoes fruits.

1.3.2. Specific objectives

 To prepare extract solution from Allium sativum.
 To use the Allium sativum extract for testing its antimicrobial properties by culturing
different microorganisms
 To preserve tomatoes using garlic extract and record weight loss at different time
point.
 everyday spice or food preservative. To collect, organize and interpret the
information generated from literature and experiments done in research project work

1.4. Research hypotheses

Use of Allium sativum (garlic) extract on Solanum lycopersicum (tomatoes) will exhibit
antimicrobial and preservative activity, leading to a reduction in microbial growth and an
increase in the shelf life of the tomatoes.
1.5. Research questions
 Is Allium sativum an effective natural preservative on tomatoes?
 Why did we choose Allium sativum as a natural food preservative?
 Are there any changes in the chemical composition or sensory characteristics of
Solanum lycopersicum (tomatoes) after the application of Allium sativum extract?
 Does the application of Allium sativum extract on Solanum lycopersicum (tomatoes)
lead to an increase in the shelf life of the tomatoes?
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1.6. Significance of study

By identifying all information about the preservative character of garlic; homes, food industries,
slaughterhouses and other food processing companies will be able to use it as a natural food
preservative. This will minimize the loss of productivity and food shortage. If the research is
successful, the information will reduce the cost and time to wait for those synthetic preservatives.
People will prevent food spoilage in their homes which will reduce the number of diseases
caused by harmful microorganisms that are likely to be in contact with food. This project will
provide a wide market which will finally result in the employment of many people. For example,
the farmers who cultivate garlic will obtain a large market for supplying their harvest. The
number of workers that industries use today will significantly increase to get enough workers to
prepare garlic for use. This research will provide other information about the health benefits of
garlic so that they can use it for other purposes than food preservation.
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CHAPTER 2. METHODOLOGY
2.1. Study area description
The samples of garlic species (both PGS and WGS) were collected in Rubavu district of Rwanda
Busasamana sector, Gihonga cell. Rubavu district yearly average temperature is 20.3°C
(https://www.weatherbase.com/weather/weathersummary.php3?s=592606&cityname=Rubavu,
+Rwanda). The average temperature for the warmest month is 20.6°C in September and July has
the coldest average temperature of any month at 19.6°C. Tomatoes were bought from
Nyabugogo modern market in Nyarugenge district of Rwanda. All the samples were brought to
the campus laboratory at University of Rwanda, College of Science and Technology located in
Nyarugenge district, Kigali city where all experiments had to be carried out. It has a tropical wet
and dry or savanna climate and is 1778.49 meters above sea level in elevation. The district
averages temperature is 21.87°C throughout the year, which is 1.43% warmer than Rwanda's
national average. The annual average in Nyarugenge brings 263.26 wet days and roughly 213.59
millimeters of precipitation (72.13% of the time).

 improve existing ones; so, old books may contain information that is outdated which can
finally give false data. The internet provides updated information which makes it more
appropriate than other sources of information.

2.3. Preparation of aqueous garlic extract

Table 2. Materials and reagents used in extraction and preparation of coating

Materials Reagents

Garlic Starch

Electric blender Distilled water

Cheesecloth Antibiotics (doxycycline)

Beaker

Centrifuge
 12

Fridge

Basin

Round bottomed flask

Centrifuge tubes

Knives
 13

Procedures

Fresh garlic bulbs were prepared according to the methodology provided by (Njue et al., 2014).
They were washed and manually separated garlic cloves. On a weighing balance, 100 grams of
peeled garlic cloves were weighed using aluminum foil. After that, 125 ml of sterile, deionized
water was added; to create a paste, the material was mixed and placed in an electric blender. The
paste was compressed using cheesecloth, and the resulting liquid was centrifuged for 10 minutes
at 5000 rpm. After that, it was transferred into a clean container and kept chilled at 4°C.

Note: For preparing the extract to be used in antimicrobial test, a sterile cheese cloth was used to
filter the mixture after clean cloves were crushed in a sterile mortar and pestle so that the filtrate
used, was regarded as a pure source of fresh garlic extract (without water).

Figure2. Sample of garlic. (A) WGS bulbs, (B) PGS bulbs, (C) WGS cloves and (D) PGS
cloves
 14

2.4. Testing antimicrobial activity of Allium sativum

Preparation of media: We chose to use nutrient agar as growth medium because it is known to
facilitate the growth of a wide range of microorganisms especially bacteria. 28 g of nutrient agar
were mixed thoroughly with 1 liter of distilled water and then sterilized in an autoclave at 121°C
for 15 minutes. It was then taken in a biosafety cabinet for cooling and poured on petri dishes.
For preparing peptone water which was used as the diluent, we mixed 15 g of peptone water
powder with 1 liter of distilled water thoroughly and sterilized at 121°C for 15 minutes and
poured 9 ml in sterile test tubes for facilitating in serial dilution.

Preparation of coating: After preparing garlic extract, we prepared laboratory starch to make a
mixture for being used as a protective coat in the ratio of starch 5:3 GE (500ml:300ml). The role
of starch is to act as a barrier for gaseous exchange. According to (Thakur et al., 2018), because
of its robust and intricate polymer network, edible coatings made from starch composites had
much better selective permeability and mechanical characteristics and may offer a way to raise
the quality of fruits.

Testing antimicrobial activity: As studies indicated, Allium sativum has general antimicrobial
activity (i.e. antifungal, antibacterial, antiparasitic and antiviral properties) (Kovarovič et al.,
2019). This means it works on a wide range of microorganisms even if it is more effective for
some of them (Rakshanaa & Geetha, 2017). For testing the antimicrobial activity of garlic
extract, we used dirty water from Happy Day Restaurant (the campus restaurant). Due to various
nutrients from food, water used for cleaning kitchen tools is likely to contain different
microorganisms (Mattick et al., 2003). So, we used it and made a serial dilution up to 10 -5 using
peptone water as diluent. We cultured on nutrient agar plates, and used agar well diffusion
method. Using a tip, we dug three holes in media on petri dishes two for positive control
(antibiotics) and negative control (the hole remained without anything) and one filled with garlic
extract in order to observe growth inhibition zone on petri dishes.
 15

2.5. Preservative activity of Allium sativum against spoilage of tomatoes

To preserve tomatoes, we used two different varieties of Allium sativum namely purple garlic
species (PGS) and white garlic species (WGS) each used to prepare edible coating separately. In
preparation of tomatoes, we used half-cut tomatoes and whole tomatoes for both varieties. We
dipped tomatoes in garlic extract for 10 minutes and then store them on shelves at room
temperature with control for both half-cut and whole tomatoes. The initial mass of all samples
were recorded followed by 3-days time interval of evaluation until their spoilage time.

2.6. Data analysis

For providing understandable and meaningful information, we used Microsoft Excel 2016 to
present data on observable charts and graphs. Microsoft excel helped to calculate the standard
deviation and mean of mass and diameter of the growth inhibition zone on petri dishes.
 16

CHAPTER 3. RESULTS
3.1. Preparation of aqueous garlic extract
The first specific aim of this study was to obtain the aqueous solution from garlic cloves. Two
garlic species: white garlic species (WGS) and purple garlic species (PGS) were selected for this
study as depicted in the figure 3A and 3B. The extracted aqueous solution presented different
colors. The extract from WGS was light yellow while that from PGS appeared as dark yellow.
Further analytical measurements showed the pH of the aqueous solution from PGS was 6. 77
while the pH of the solution extracted from WGS was 6.92.

Figure 3:
Preparation of garlic aqueous solution. Two varieties: white garlic cloves(A) and purple garlic
 17

cloves (B) were selected and used to produce white garlic solution (E) and purple garlic solution
(F).

3.2. In vitro antimicrobial properties

The antimicrobial activity of aqueous solution extracted from garlic was tested against
microorganisms from dirty water collected from Happy Day Restaurant sinks using agar well
diffusion method. WGS, PGS alongside with negative and the antibiotic as the positive control
were inoculated on the plates (see figure 4). The growth inhibition zone was calculated by
measuring the diameter of circle that surrounds a space where microorganisms did not grow due
to the antimicrobial activity. The results of the zone of inhibition for all group samples are
demonstrated in the table 5.

Figure 4. Antimicrobial activity. The growth inhibition zone due to antimicrobial activity for
PGS & WGS with their respective positive and negative control.
 18

Table 3: Variation of growth inhibition zone

Serial dilutions for both WGS Inhibition zone by extract Inhibition zone by antibiotics
and PGS (cm) (cm)
WGS (100) 1.7 2.1
WGS (10-1) 1.5 3
WGS (10-2) 1.5 3
WGS (10-3) 1.9 3.4
WGS (10-4) 2 4.5
WGS (10-5) 2 4
PGS (100) 1.4 2.4
PGS (10-1) 1.7 2.7
PGS (10-2) 1.9 3
PGS (10-3) 2.2 4
PGS (10-4) 2.8 4.2
PGS (10-5) 2.7 4.5

Figure 5. Graph represent the length of growth inhibition zone

 19

3.3. Preservative activity of Allium sativum on fresh tomatoes.

As illustrate in the table 7, after dipping tomatoes (both half-cut and whole tomatoes) in garlic
extract for 10 min, the mass of tomatoes was recorded. It is observable that the mass was
decreasing with time and spoilage of uncoated tomatoes started appearing at day 6 for half-cut
and their smell started to change (changing to bad smell) while fresh tomatoes have a grassy
smell. At day 9, the half-cut ones were completely spoiled and further investigation could not be
carried out. On the figure 5, we can see that on day 6 there were some microorganisms grew out
on uncoated tomatoes; however, coated tomatoes were not affected at the same extent as
uncoated tomatoes.

Figure 6: Preservative activity of garlic on half cut tomatoes. The number in brackets stands
for days of preservation. PGS & WGS stands for purple garlic species and white garlic species
respectively.
 20

If we look at the figure7, on day 9 we observe that all tomatoes were spoiled but the uncoated
appeared to spoil before and more than the others. The smell was then worse and the tomatoes
appeared to be exhausted with flaccid and dry tissue.

Figure 7: Preservative activity of garlic on tomatoes at day 9.

On the other hand, whole tomatoes appeared to spoil after relatively long time compared to the
half-cut tomatoes. Physical appearance started to change on day 12, where they started wilting as
as result of dehydration. Furthermore, the rate at which mass of whole tomatoes decreases is low
compared to that of half-cut ones.

Figure 8: Preservative activity of garlic on whole tomatoes. Image of coated and uncoated
tomatoes for first 6 days and last 9 days. The number in bracket stands for the preservation day.
 21

The table below contains data on weight loss of tomatoes at different time point after
preservation followed by statistical descriptive graphs indicating percentage weight loss for half-
cut tomatoes (figure 9) and whole tomatoes (figure 10).

Table 4: Decrease in a mass of preserved tomatoes over time and their corresponding
standard deviation.

Days Mas Mas Mas

Mass s s s Mass Mass Mass Mass Mass Mass
Day0 Day Day Day Day1 Day1 Day17 Day2 Day2 Day2
(gr) 3 6 9 2 (gr) 5 (gr) (gr) 1 (gr) 4 (gr) 7 (gr)
Samples (gr) (gr) (gr)
Whole
165. 163. 159. 155.
tomatoes 149.8 147.9 146.2 143.4 140.5 136.9
6 1 7 4
(WGS)
Whole
105. 104. 102. 100.
tomatoes 97.6 95.2 93.5 90.8 88.9 86.8
7 3 7 3
(PGS)
Whole
123. 121. 118. 116.
tomatoes 113.2 111.2 109.9 107.8 105.6 102.7
4 7 7 7
(non-coated)
Half-cut
97.5 87.2 73.6 55
(PGS)
Half-cut
45.8 38.2 28.4 16.8
(WGS)
Half-cut
69.7 59.8 46.1 27.7
(non-coated)
 22

Figure 9: Weight loss percentage for half cut tomatoes

Figure 10: Weight loss percentage for whole tomatoes

 Non-cut tomatoes for both uncoated and preserved ones, it seems that in the first days of
preservation, there is little difference in mass loss (i.e. the mean mass lost for tomatoes
preserved by WGS, PGS and uncoated were 3.17, 2.12 and 2.22 respectively).
 23

Even if there is no big difference in mass lost, it can be observed that whole tomatoes
preserved by PGS maintained mass than those preserved by WGS with (P<4.36*10 -12).
However, the whole tomatoes reached at day 21 without observable decay; the slight
change of external color is due to wilting which appeared more on uncoated tomatoes and
tomatoes preserved by PGS appear to be fresh compared to those preserved by WGS. The
weight loss of preserved half-cut tomatoes were different with that of non-preserved with
(P< 2.22*10-6) for uncoated and those coated by PGS and (P<0.016) for uncoated and
those preserved by WGS. However, according to (Desta Dugassa, 2021), weight losses
differed significantly (P<0.05) starting on day 6 of storage (4.26% for the control, 2.79%
for the starch and garlic coatings), and the largest weight loss was 9.06% on day 30 for
tomato fruits coated with maize starch. The control sample, on the other hand, showed
consistently (P<0.05) the largest percentage of losses over the course of the storage
period, with the highest loss occurring on day 30 (14.71%).
 24

CHAPTER 4. DISCUSSIONS
4.1 Preparation of garlic aqueous solution
 Yield: In preparation of garlic extract we used 376gr for WGS and 860gr for PGS which
generated 400 milliliter (ml) and 900 milliliter (ml) respectively and this yield seems not
to be different according to mass used. However, (Klinger, 2020) said that fresh purple
garlic has a milder flavor and is "juicier" than white garlic.
 Nutritional value: According to (WENLI, 2022), protein, vitamins, fiber, phosphorus,
iron, calcium, and other mineral components are all present in garlic. Modern medical
investigation revealed that purple garlic is nearly identical to white garlic in terms of
nutrients, with the exception of anthocyanin (whose main function is an antioxidant).
Additionally, scientists have discovered that the antibacterial properties of purple-skinned
garlic are more prominent and its allicin content is relatively high.
 Difference in colors of extracts: The extract from WGS was light yellow while that
from PGS appeared as dark yellow. According to (Imai et al., 2006), the enzymatic
interactions between pigment precursors (i.e. anthocyanin as said by Fossen & Andersen,
(1997)) , amino acids, and allicin give garlic extract determine the final color. These
reactions happen as a result of the fragmentation of the garlic and the release of the
enzyme allinase, which converts alliin to allicin. However, by employing heat as a
catalyst and color developers to produce the color of garlic juice, such reactions can be
carried out artificially. Therefore, the distinct colors of the PGS and WGS extracts may
be caused by their content in various pigment precursors.
 Antimicrobial activity
After 24 hours of incubation at 37°C, the petri dishes indicate that antibiotics showed a
wider growth inhibition zone than that of both PGS and WGS (see figure4). However,
the growth inhibition zone caused by PGS seems to exceed that of WGS according to the
mean length they provided. For example, let’s take two serial dilutions i.e. 10 -1 and 10-2
for both varieties; WGS had 1.5 for both dilutions while PGS had 1.7 and 1.9
respectively. On another side, antibiotics on WGS plates had 3 cm on both dilutions
while on PGS plates had 2.7 cm and 3cm respectively. The mean lengths of the growth
inhibition zone (diameter) provided by antibiotics, PGS and WGS are 3.4cm, 2.11cm and
1.7cm, respectively. Literatures has been thoroughly shown in vitro and in food systems
 25

that spices and garlic have an inhibitory effect against a wide range of microorganisms,
including viruses, bacteria, fungus, and parasitic protozoa with mean diameter of 4.2 (±
0.6) for garlic. Yet, depending on geographic locations, seasons, and environmental
factors, the spectrum of inhibition varies widely amongst spices and even within the same
spice (Benkerroum & Patel, 2018).
From the study carried out by (El-Sayed et al., 2017), the following were observed:
Compounds Diallyl Trisulfides and Diallyl Disulfides made up, respectively, 0.36% and
2.51% of WGS, and 0.71% and 3.16% of PGS. This results in the composition of WGS
and PGS containing, respectively, 64.26% and 84.78% of the total diallyl sulfides. This
research revealed that the antibacterial properties of garlic essential oil (GEO) are due to
the disulfide bonds (S–S) of its constituent parts. As a result, the antibacterial activity
increases in the following order as the number of disulfide links in the component
increases: tetrasulfide > trisulfide > disulfide > monosulfide. If one were to apply those
characteristics to this study, one may predict that PGS would exhibit greater antibacterial
activity than WGS. Especially at higher concentrations, WGS demonstrated greater
antibacterial efficacy than PGS against the studied pathogens. It appears that this is
because WGS (70.03%) has a greater percentage of total trisulfides, which may
somewhat offset PGS's higher percentage of total disulfides. Only one disulfide bond (1
S-S) was present in the disulfide molecules, compared to two disulfide bonds (2 S-S) in
the trisulfides, which had greater WGS concentrations and stronger antibacterial action.
 Weight loss: According to (Desta Dugassa, 2021), the percentage weight loss (%w) for

each sample was calculated by the following formula: %W= where Wi is

initial mass (on the first day) and Wf is the final mass (on last day). If we look at (figure
8), we can see that tomatoes preserved by PGS retained mass more than uncoated
tomatoes and those preserved by WGS. On the other hand, in the first 6 days, tomatoes
preserved by WGS showed higher weight loss than uncoated tomatoes even if the
following 3 days it lost weight to a higher extent because its peak approached that of
WGS. It is obvious that if we had the chance to reach on day 12, the peak of control
would be exceeding that of WGS.
 26

 Smell: The smell of fresh tomatoes seems to be grassy. The most potent aldehyde to
contribute to the olfactory profile of the cherry tomato is (Z)-3-hexenal, which produced
a green-grassy odor. It was followed by (E)-2-hexenal (green, leafy), and hexanal (fresh,
cut grass) (Selli et al., 2014). They stated that fresh tomatoes have a flavor that comes
from a complex interaction of sugars, organic acids, volatile compounds made of
carotenoids like lycopene and beta carotene and amino acids during fruit ripening. Other
volatile compounds are created when lipids break down in tissues that have been
damaged, forming primarily C6 compounds through enzyme cascade. When the spoilage
starts, the smell changes and the odor become fusty as time goes especially for half-cut
tomatoes. We can absolutely state that bad smell increases as the time of spoilage is
prolonged.
 Spoilage: As we have discussed in literatures review, tomato microorganisms spoilage is
enhanced by natural rotting by pectolytic enzymes polygalacturonase (PG), pectin
methylesterase (PME), and pectate lyase (PEL) which degrade pectin in tomato’s cell
wall (Rosales et al., 2009). This contribute to its softening and finally support
microorganisms attack. According to (Skrzypiec, 2022), Tomatoes that are not yet ripe
require two to seven days to mature. Cherry tomatoes keep their quality for around 4 to 6
days at room temperature or up to two weeks in the refrigerator once they are ripe, while
grape tomatoes last for about 3 to 4 days on the counter or up to 10 days in the fridge.
However, the length of time they spend on the seller's shelves, the storage environment's
temperature, and the storage conditions can all affect that time. On the other hand, as the
figure 8 shows, on day 21 tomatoes were still good to be used which is contrast to the
time provided by (Skrzypiec, 2022). This indicate that garlic extract inhibited growth of
microorganisms that spoil tomatoes and starch reduced the rate of gaseous exchange,
which prolonged their shelf life from 6 to 27 days and can be the sign of preservative
activity of garlic (Allium sativum). Still and all, this may also be due to the climate of
where experiment took place, cultivation methods (pectin content and pectolytic enzymes
activity due to type of soil, fertilizers or pesticides used), harvesting, packaging,
transportation and storage conditions such exposure to various microorganisms.
As shown in the figure 6, there were microorganisms growth on tomatoes. In his study,
(Ghosh, 2009) came to the conclusion that Aspergillus niger, Fusarium, and Penicillium
 27

were the main fungus responsible for the tomato fruit's destruction. Additionally, these
fungi are a source of strong mycotoxins like ochratoxin, which is produced by A. niger
and is likely carcinogenic, causing esophageal cancer and endemic nephrotoxicity in the
Balkans, and trichothecenes, which is produced by fusarium and exhibits a wide range of
diseases, some of which are even fatal. Thus, rotten tomatoes should never be ingested
and should never be fed to livestock. (Sidney, 2018) showed that Molds begin as airborne
spores that travel across the environment. When these spores land on spoiled food, they
germinate and produce a mass of filamentous hyphae that eat their way through the
spoiled food and develop into the vague, whitish mass known as the mycelium.
Ultimately, from the mycelium, specialized hyphae emerge to form upright stalks, on top
of which fruiting bodies develop (sporangia). The mature sporangia of this genus are dark
grey. The sporangial wall eventually breaks down, releasing fresh spores into the
atmosphere.

Proposed limitations of the study

 Collection of tomatoes: normally, tomatoes don’t ripe at the same time. One can ripe
today while another can ripen tomorrow and so on; however, they are all carried to the
market at the same time. When we go to the market to buy tomatoes, we try to choose
tomatoes that seem to be at the same level of ripening but we are not sure about the
precision of that choice. Tomatoes can also be collected from different regions with
different soil, cultivation methods, harvesting methods, packaging, transportation and
storage conditions that can influence the shelf life of tomatoes whether or not they are
preserved.
 Collection of garlic species: the same case here, cultivation conditions and region where
Allium sativum is grown matters to the concentration of chemical compounds. Also the
amount of solvent used during extraction can affect the concentration of bioactive
compounds found in garlic extract. So, it is better to know whether to use crude juice or
dilute juice.
 28

 Collection of wastewater for anti-microbial test: while collecting wastewater, we had

to make sure it is dirty enough and not hot to contain alive microorganisms. Because we
were testing general antimicrobial activity, we had to make sure the source contains
nutrients that allow them to grow and replicate.
 29

CHAPTER 5. CONCLUSION AND RECOMMENDATIONS

5.1: Conclusion
We can confirm that Allium sativum has antimicrobial activity. However, this activity is
determined by the concentration of garlic in the coating. The fact that garlic can be mixed with
other constituents i.e. starch is because they act as a supporting element such as making edible
coating elastic, semipermeable and so on when coated to the fruits. As the variation of mass loss
and growth inhibition zone against microorganisms, we can conclude that the purple garlic
varieties are more effective than the white varieties (Kay, 2023). So, in order to minimize fruits
and vegetables productivity loss and keep them safe when brought to home, we can use garlic
based edible coatings to minimize the post harvest loss.

5.2: Recommendations
 Recommendation to researchers: we recommend all researchers who are studying
garlic as a food preservative (or who will be interested in this field) to work on the proper
and effective concentrations of garlic which can be used in edible coatings to be used in
the preservation of fruits and vegetables. They can also look at other natural ingredients
which will improve the effectiveness of garlic in preservation. This will make coating
nontoxic, elastic, and improve the nutritional and pharmaceutical properties of vegetables
and fruits.
 Recommendation to UR-CST: We recommend upgrading laboratory materials so that
students can get all requirements for the proposed project. We also recommend UR-CST
to support students' projects financially so that their ideas cannot be limited by financial
issues.
 Recommendation to garlic consumers: we have seen that Allium sativum (garlic) has
antimicrobial and preservative activity. By identifying this information about the
preservative character of garlic; homes, food industries, slaughterhouses and other food
processing companies might use garlic as a natural food preservative. This will minimize
the loss of productivity and food shortage. Moreover, this is a call for the use of natural
edible coating instead ofsynthetic preservatives. People will prevent food spoilage in
their homes which will reduce the number of diseases caused by harmful microorganisms
that are likely to be in contact with food.
 30

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APPENDIX
The 17 aminoacids that are found in Allium sativum (garlic)

 Cysteine  Tryptophan
 Aniline  Phenylalanine
 Isoleucine  Arginine
 Threonine  Proline
 Serine  Leucine
 Aspartic acid  Lysine
 Histidine  Tyrosine
 Glutamic acid

 Valine

Figure 11. The chemical structure of alliin, allicin and its derived OSCS

Cultivation of Allium sativum (garlic)

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1. Crop rotation and location

Rotation is a useful method to use when growing garlic. Never cultivate garlic where onions or
another member of the onion family has previously been grown. Garlic should be planted in full
sun and on a bed with organic matter incorporated that is well-drained(Wayne J. McLaurin,
Adams, David, 2015).

2. Planting

Cloves, the tiny bulblets or segments that make up the garlic bulb, which are planted instead of
the actual seed that garlic produces. Dig a hole, insert the unpeeled clove carefully in the hole
with the pointy side up, and then cover the clove with soil to properly plant garlic. A straight
neck is ensured by placing the cloves upright. Note: Divide the bulb only when you are ready to
plant it because early separation reduces yields. Choose "seed bulbs" that are big, smooth,
healthy, and disease-free.

After planting, garlic cloves need 6–8 weeks of cool weather to go through vernalization, which
is the process of being forced to form a bulb and bloom. The plant enters a phase of vegetative
development as soon as there is enough moisture and a decrease in temperature. Once bulb
formation starts, don't fertilize again. Maturity is indicated when leaves start to turn brown and
tops start to fall. To prevent bulb rot and discoloration, stop irrigation at this time. Remove the
top layer of dirt from a few bulbs and examine them to make sure they are totally distinguishable
to make sure the bulbs are fully grown. Didigging premature bulbs, can result in bulb
deterioration, disease and discoloration during long time storage (Wayne J. McLaurin, Adams,
David, 2015).

3. Storage

The secret to keeping your garlic fresh is to store it in a dark place where the temperature doesn't
change drastically and it gets enough air circulation. For storage over the winter, any cool, well-
ventilated area would do. The bulbs should be kept from freezing in extremely cold climates.
The best circumstance is to keep a mesh bag full of garlic in a seldom-used cabinet or cool cellar.
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There should be about 70% humidity. Make sure the remaining bulbs of garlic are not exposed to
light when you remove them for use. This will prevent the bulbs from going into growth and
shorten the amount of time they will remain fresh.

Tomato postharvest loss around the world

According to (Arah et al., 2015)he main causes of tomato postharvest loss include: inappropriate
harvesting periods, lack of appropriate harvesting containers, excessive field heats & lack of on-
farm storage facilities, inappropriate packaging materials and poor Field Sanitation.

Figure 12. Image showing global postharvest loss of tomatoes