Development of Safe Juices For Immunocompromised Patients by Irradiation Alone or in Combination With Other Technologies

FacultyofScience
Microbiologydepartment
Development of safe juices for

immunocompromised patients by irradiation
alone or in combination with other
technologies
By
Reham Mohamed Mohamed Abd El-kader

B. Sc. Microbiology Biochemistry
(2007)
Faculty of Science, Ain- shams University
National Center For Radiation Research and Technology
Thesis
submitted in partial fulfillment of the requirement for
M. Sc. Degree in
Microbiology
Microbiology Department
Faculty of Science
Ain-Shams University
2012
FacultyofScience

technologies
By
Reham Mohamed Mohamed Abd El-Kader

B. Sc. Microbiology - Biochemistry
(2007)
Supervisedby:
Prof. Khayria Abd El-Ghany Youssef

Professor of Microbiology, Faculty of Science, Ain Shams
University
Prof. Ali Ahmed Ibrahim Hammad

Professor of Food Science and Microbiology, FormerVice
Chairman of Egyptian Atomic Energy Authority
Dr. Hanan Hassan Abd El-Khalek

Lecturer of Microbiology, National Center for Radiation
Research and Technology (NCRRT), Atomic Energy Authority
2012

FacultyofScience
Name: Reham Mohamed Mohamed Abd El-Kader

Title:
Development of safe juices for immunocompromised
patients by irradiation alone or in combination
with other technologies
Supervisedby:
1-Prof. Khayria Abd El-Ghany Youssef

ProfessorofMicrobiology,FacultyofScience,AinShamsUniversity.
2-Prof. Ali Ahmed Ibrahim Hammad..

ProfessorofFoodScienceandMicrobiology,FormerViceChairmanofEgyptian
AtomicEnergyAuthority.
3- Dr. Hanan Hassan Abd El-Khalek.

LecturerofMicrobiology,NationalCenterforRadiationResearchandTechnology
(NCRRT),AtomicEnergyAuthority(AEA).

FacultyofScience

technologies
By
Reham Mohamed Mohamed Abd El-Kader

B. Sc. Microbiology - Biochemistry
(2007)
Approvedby:
1-Prof. Mohie Eddin Zohier El-Fouly

Professor of Microbiology, National Center For Radiation
Research and Technology ( NCRRT ), Atomic Energy Authority
2-Prof. Mohamed Ghareeb Ibrahim Emaish

Professor of Microbiology, Faculty of Education, Ain Shams
University.
3-Prof. Khayria Abd El-Ghany Youssef

Professor of Microbiology, Faculty of Science, Ain Shams
University
4-Prof. Ali Ahmed Ibrahim Hammad

Professor of Food Science and Microbiology, FormerVice
Chairman of Egyptian Atomic Energy Authority
DEDICATION
To my mum Mrs.Nawal Mahmoud.
Your constant motivation and faith has always given
me hope. Now, this is was your dream, through me.
Thank You.
&
To my brother Islam Abdelkader for his supports
and encouragements.
D
&
To my father soul, God bless him.
Finally, this thesis is dedicated to all those who believe
in the richness of learning.
ACKNOWLEDGEMENTS
All praises and thanks are for ALLAH who is the
entire source of all knowledge and wisdom endowed
to mankind
Prof. Khayria Abd El-Ghany Youssef, thank you for believing in me; for
your encouragement through kind words, support, your guidance, for
effective and helpful supervision, Fruitful discussion and valuable
advice.
Prof. Ali Ahmed Ibrahim Hammad, thank you for suggestion of the point,
assigning the work, kindly supplied me with all necessary facilities,
carefully editing this thesis and your attention to finer details, and
thanks for your encouragement, guidance, patience, time and expertise on
this work.
My heartfelt gratitude goes to Dr. Hanan Hassan Abd El-Khalek for all
her help, encouragement and dedication in helping me, support, patience,
and sincere advices, advance my knowledge and making this work
possible.
Prof Hussein Abd El-Kareem Head of Microbiology department, National
Center for Radiation Research and Technology (NCRRT).Atomic Energy
Authority(AEA).A cooperative and affectionate personality, special
thanks are extended to you for the care, impetus and encouragements.
Thanks to the head of Microbiology department Mohamed khaled
Ibrahim and to the faculty, staff and members of the Microbiology
department for their help and their academic advice.
My deepest graditude to all staff of Microbiology department,

National Center for Radiation Research and Technology for their
friendship and their advice.
Last, but not least my deep thanks are extended to my family and all my
friends, their love, understanding and unfailing support has made
everything possible.
I thank all those who, in different ways, have walked beside me along
the way; offering support and encouragement, challenging my thinking,
and teaching me to consider alternative views.
Name : Reham Mohamed Mohamed Abd El-Kader

Title of Thesis : Development of safe juices for immunocompromised patients by
irradiation alone or in combination with other technologies.
Degree: M.Sc.
Faculty : Science
University : Ain Shams
Department : Microbiology
Abstract
Fresh fruit and vegetable juices have become a new functional food
available for dietary and health. However, they have high microbial load and poses a
microbial hazard to consumer unless treated with a suitable method of food
preservation. Gamma irradiation, as non thermal process, is highly effective in
destruction of both spoilage and pathogenic microorganism in various foods without
arising its temperature, hence keeping its freshness and nutritional quality. In recent
years, it is used as a safe and alternative method for food preservation in many
countries.
In present study, pomegranate, tomato and carrot juices were irradiated at
different irradiation doses (1- 4.5 kGy). Yeasts, as the principle cause of juices
spoilage, were isolated and identified from unirradiated (control) juices. The
resistance of the identified yeasts to gamma radiation in the term of D10- value was
investigated. It was found that Debaryomyces hansenii had the highest radiation
resistance ( D10-value of 1.63 kGy ) while Candida tropicalis had the lowest
resistance to gamma radiation ( D10-value of 0.98 kGy ).The effect of these
irradiation doses on the microbiological load, nutritional, physiochemical and
sensory properties were evaluated immediately after irradiation and during
subsequent refrigeration storage. Generally, irradiation doses of 1.0 kGy and above
were efficient and sufficient for complete elimination of coliform bacteria and E.coli
found in these juices without impairing the nutritional and sensory quality attributes.
Irradiation of pomegranate juice at 2.5 kGy greatly reduced its microbial
counts and extended the shelf-life to 14 days at refrigeration temperature (4C1)
with minimal changes in the anthocyanin content. Irradiation dose of 3.0 kGy could
improve the microbial quality, ensure safety and extend the shelf-life of tomato juice
to 15 days at refrigeration temperature without significant changes in lycopoene
content. Meanwhile, irradiation dose of 3.0 kGy in combination with
cinnamaldehyde (8l/100 ml) greatly reduced the microbial load of carrot juice and
extended its shelf-life to 21 days at 4C1 without adverse effect on the sensory and
nutritional quality.
Key words : pomegranate juice, tomato juice, carrot juice, irradiation, nutritional
quality.
Contents
Contents
Page
Introduction
Review of Literature
1. Immunocompromised patients
2. Immunocompromised patients problems
3. Immunocompromised patients recommended diets
4.Important of fresh fruits and vegetables and their juices for human
health
10
4.1. Pomegranate juice
13
4.2. Tomato juice
14
4.3. Carrot juice
15
4.4. Anthocyanin
15
4.5. Lycopene
16
4.6. Carotenoids
18
4.7. Ascorbic acid
19
5. Microbial contamination of food in hospitals
20
6. Microbial contamination of fresh fruits and vegetables
23
7. Microbial contamination of fresh fruits and vegetables juices
25
8. Methods used to reduce the hazards of foodborne infection for

immunocompromised patients
27
8.1. Thermal treatments
27
8.2. Refrigeration
29
8.3. Chemical additives
30
Contents
8.4. High hydrostatic pressure processing ( HHP )
30
8.5. Ionizing radiation
31
9. Treatment of food by irradiation
32
9.1. Radiation sources used for food irradiation
33
9.2. Wholesomeness and safety aspects
33
9.3. International approval
34
9.4. Technological benefits and advantages
36
9.5. Limitations
37
10. Irradiation effects
38
10.1. Relative radiation resistance of microorganisms
39
10.2. Effect of irradiation on microbial load of fresh

fruits and vegetables
41
10.3. Effect of irradiation on foodborne pathogenic

bacteria contaminating fresh fruits and vegetables
and their minimally processed produce
42
10.4. Effect of irradiation on microbial quality of fruits

and vegetables juices
44
10.5. Effect of irradiation of the main chemical

components of fresh fruits and vegetables and their
produce
45
10.5.1. Anthocyanin
45
10.5.2. Lycopene
47
10.5.3. Carotenoids
47
10.5.4. Ascorbic acid
48
10.6. Effect of irradiation on sensory properties
50
10.7. Viscosity effect
52
11. Preservation of juices by irradiation and other

technologies
11.1. Irradiation in combination with refrigeration
II
53
54
Contents
11.2. Irradiation in combination with natural additives
56
Materials and Methods
59
1. Preparation of Juices
59
1.1. Preparation of fresh squeezed pomegranate juice
59
1.2. Preparation of fresh squeezed tomato juice
59
1.3. Preparation of fresh squeezed carrot juice
60
2. Gamma irradiation
60
3. Storage
61
4. Microbiological analysis
61
4.1. Total aerobic bacterial counts ( TBC)
61
4.2. Lactic acid bacteria ( LAB)
61
4.3. Total molds and yeasts
62
4.4. Total coliform
62
4.5. Escherichia coli
62
4.6. Enterococcus faecalis
64
4.7. Staphylococcus aureus
64
4.8. Aeromonas hydrophila
65
5. Isolation and purification of yeasts
65
6. Identification
66
6.1. Identification of yeast isolates
66
6.2. Incubation temperature
66
6.3. Microscopical examination
66
6.4. Biochemical tests
66
6.5. Urease test
67
6.6. Cycloheximide sensitivity test
67
7. Determination of D10 Values
67
7.1. Preparation of inoculum
68
III
Contents
7.2.Inoculation
68
7.3. Irradiation
68
7.4.Plating
68
7.5. D10 - value
69
8. Nutritional analysis
69
8.1. Anthocyanin content in pomegranate juice
69
8.2. Lycopene content in tomato juice
70
8.3. Total carotenoids in carrot juice
71
8.4. Ascorbic acid measurment
71
9. Physiochemical analysis
72
9.1. Viscosity
72
9.2. pH
72
10.Sensory evalutation
72
11. Combination treatments
73
12. Statistical analysis
74
Media and reagents used in Microbiological analysis
74
Results and Discussion
84
1. Isolation and Identification of yeasts
84
2. Radiation-decimal reduction dose ( D10- value )
90
3. Irradiation of fresh pomegranate juice
99
3.1. Effect of - irradiation and refrigeration storage on the microbial

load of pomegranate juice
99
3.2. Effect of - irradiation and refrigeration storage on the pathogenic

bacteria contaminating pomegranate juice
102
3.3. Effect of - irradiation and refrigeration storage on the anthocyanin

content of pomegranate juice
1044
IV
Contents
3.4. Effect of -irradiation and refrigeration storage on

physiochemical properties of pomegranate juice
a) Viscosity
107
b) pH
109

sensory evaluation of pomegranate juice
4. Irradiation of fresh tomato juice
111
115
4.1. Effect of - irradiation and refrigeration storage on the

microbial load of tomato juice
115

pathogenic bacteria contaminating tomato juice
118

nutritional quality of tomato juice
120
a- ascorbic acid content
120
b- lycopene content
122

physiochemical properties of tomato juice
124
a) Viscosity
124
b) pH
126

sensory evaluation of tomato juice
5. Irradiation of fresh carrot juice
128
132

microbial load of carrot juice
132

pathogenic bacteria contaminating carrot juice
135
5.3. Effect of - irradiation and refrigeration storage on

the nutritional quality of carrot juice
138
a- ascorbic acid content
138
b- total carotenoids content
140
107
Contents

physiochemical properties of carrot juice
143
a) Viscosity
143
b) pH
144

sensory evaluation of carrot juice
146
6. Combination treatment of irradiation and cinnamaldehyde
149
6.1. Effect of irradiation and cinnamaldehyde on the

149
6.2. Effect of irradiation and cinnamaldehyde on

152

155
a) Viscosity
155
b) pH
156

158
Summary and Conclusion
160
References
170
Arabic Summary
VI
List of Tables
Lists of Tables
Page
Table 1: The approximate content of lycopene in different fruits
and vegetables and other foods
Table 2 : French recommended microbiological limits for
different items of food
Table 3 : Yeast identification
Table 4: Identification of yeasts isolated from juices
Table 5 : Effect of incremental irradiation doses on the viable
counts ( cfu. ml-1) of Rhodotorula glutinis
counts ( cfu. ml-1) of Candida tropicalis
counts ( cfu. ml-1) of Trichosporon pullulans
counts ( cfu. ml-1) of Debaryomyces hansenii
counts ( cfu. ml-1) of Saccharomyces cerevisiae
Table 10: Effect of -irradiation and subsequent storage ( 4C )
on the microbial counts ( log cfu/ml ) contaminating
pomegranate juice
on the some pathogens contaminating pomegranate
juice
Table 12: Total anthocyanin content in pomegranate juice
during storage at 4C1
Table 13: Viscosity (cPs) of pomegranate juices treated by
irradiation during storage at 4C1
Table 14 : pH value of irradiated and non-irradiated
pomegranate juices during storage at 4C1
Table 15: Effects of irradiation on the sensory scores of
pomegranate juice during storage at 4C1
VII
18
21
86
87
92
93
94
95
96
100
103
105
108
110
112
List of Tables

tomato juice
Table 17 : Effect of -irradiation and subsequent storage ( 4C )
on the some pathogens contaminating tomato juice
Table 18 : Ascorbic acid content ( mg/100 ml ) of tomato juice
treated with irradiation during storage at 4C1
Table 19 : Lycopene content in irradiated and non-irradiated
tomato juice ( mg/100 g wet weight ) during storage at
4C1
Table 20: Viscosity (cPs) of tomato juices treated by irradiation
Table 21 : pH value of irradiated and non-irradiated tomato
juices during storage at 4C1
Table 22: Effects of irradiation on the sensory scores of tomato
juice during storage at 4C1
carrot juice
on the some pathogens contaminating carrot juice
Table 25 : Ascorbic acid content ( mg/100 ml ) of carrot juice
Table 26 : Effect of irradiation and refrigeration storage on the
total carotenoids of carrot juice
Table 27: Viscosity (cPs) of carrot juices treated by irradiation
Table 28 : pH value of irradiated and non-irradiated carrot juices
Table 29 : Effects of irradiation on the sensory scores of carrot
VIII
116
119
121
123
125
127
130
133
137
139
142
143
145
147
List of Tables
Table 30 : Effect of irradiation and cinnamaldehyde on the

microbial counts ( log cfu/ml ) contaminating carrot
Table 31 : Effect of irradiation and cinnamaldehyde on
pathogens contaminating carrot juice during storage at
4C1
Table 32: Viscosity ( cPs) of carrot juice treated by irradiation
in combination with cinnamaldehyde
Table 33: pH values of carrot juices treated by irradiation in
combination with cinnamaldehdye
Table 34: Effect of irradiation and cinnamaldehyde on
sensory evaluation of carrot juice during storage at
4C1
IX
151
154
155
157
159
List of Figures
Lists of Figures
Fig. 1 : Carotenoid profile of tomato products
Fig. 2 : Structure of Anthocyanin
Fig. 3 : Structure of all-trans lycopene
Fig. 4 : Structure of -carotene
Fig. 5 : Structure of ascorbic acid
Fig. 6: Effect of incremental irradiation doses on the viable
counts ( cfu. ml-1) of Rhodotorula glutinis in carrot
juice
counts ( cfu. ml-1) of Candida tropicalis in
pomegranate juice
counts ( cfu. ml-1) of Trichosporon pullulans in
tomato juice
counts ( cfu. ml-1) of Debaryomyces hansenii in
pomegranate juice
Fig. 10 : Effect of incremental irradiation doses on the viable
counts ( cfu. ml-1) of Saccharomyces cerevisiae in
tomato juice
Fig. 11 : Total anthocyanin content in pomegranate juice
Fig. 12 : Viscosity (cPs) of pomegranate juices treated by
Fig. 13 : pH value of irradiated and non-irradiated pomegranate
Fig. 14 : Effects of irradiation on the sensory scores of
pomegranate juice during storage at 4C1
Page
14
16
17
19
20
92
93
94
95
96
106
108
110
113
List of Figures
Fig. 15 : Ascorbic acid content ( mg/100 ml ) of tomato juice

Fig. 16 : Lycopene content in irradiated and non-irradiated
tomato juice ( mg/100 g wet weight )during storage at
4C1
Fig. 17 : Viscosity (cPs) of tomato juices treated by irradiation
Fig. 18 : pH value of irradiated and non-irradiated tomato
Fig. 19 : Effects of irradiation on the sensory scores of tomato
Fig. 20 : Ascorbic acid content ( mg/100 ml ) of carrot juice
Fig. 21: Effect of irradiation and refrigeration storage on the
total carotenoids of carrot juice
Fig. 22 : Viscosity (cPs) of carrot juices treated by irradiation
Fig. 23: pH value of irradiated and non-irradiated carrot juices
during storage at at 4C1
Fig. 24 : Effects of irradiation on the sensory scores of carrot
Fig. 25: Viscosity ( cPs) of carrot juice treated by irradiation
in combination with cinnamaldehyde
Fig. 26: pH values of carrot juices treated by irradiation in
combination with cinnamaldehdye
Photos :
Photo 1 : Candida tropicalis
Photo 2 : Debaryomyces hansenii
Photo 3: Saccharomyces cerevisiae
Photo 4 : Trichosporon pullulans
Photo 5 : Rhodotorula glutinis
XI
122
124
126
127
131
140
142
144
145
148
156
157
87
88
88
89
89
Abbreviations
Abbreviations
AD Alzheimers disease
AOACAssociation of Official Analytical Chemists.
APHA American Public Health Association
ATBC drinks alpha-tocopherol-beta carotene drinks
AwWater activity
BMTBone marrow transplantation
BPA Baired-Parker Agar
cfucolony forming units
ETsEllagitannins
FAO Food and Agriculture Organization
HHP High hydrostatic pressure
HIV Immunodeficiency virus
HMFHydroxymethyl furfural
IAEAInternational Atomic Energy Agency
IARC International Association of Research on Cancer
ICGFI International Consultative Group on Food
Irradiation
ICH Immunocompromised host
ICMSF International Commission on Microbiology
specifications for food
IFT Institute of food technologies
KAAAKanamycin Aesculin Azid Agar
LABLactic acid bacteria
LBDs Low bacterial diets
XII
Abbreviations
MPEDMost probable effective dose

MPNMost Probable number
MRSMan, Rogosa and Sharp agar
PGPoly galacturonase
PME Pectin methylesterase
RMPFRefrigerated minimally processed foods
RTEF Ready to eat foods
SAA Stach ampicillin agar medium
TBCTotal aerobic bacterial count
TM&YTotal mold and yeasts
USFDAUnited States of Food and Drug
Administration
WHO World Health Organization
YCB Yeast carbon base
YNBYeast nitrogen base
NCRRT National Central for Radiation Research and
Technology
CpsCentipoise
D10The radiation decimal reduction dose
kGy kilogray
XIII
Introduction
Introduction
The immunocompromised host (ICH) has impaired
defense mechanisms, rendering the patient more susceptible to
infection. The immune system is primarily disrupted by
impaired function of T cells, or neutrophils. By definition, the
immunocompromised state results from impairment of the host
defenses; thus, conditions affecting the function and number of
neutrophils, macrophages, lymphocytes, and circulating
antibodies predispose the host to certain infections (Safadi and
Soubani, 2009). The most common defect in neutrophils is a
decrease in their absolute number, as is characteristic of patients
with leukemia, Aplastic anemia, or those who have received
radiation or chemotherapy. Infectious diseases represent a
major cause of morbidity and mortality in the patient with
compromise in immune function (Young and Rubin, 1994).
There is an increasing recognition that immunocompromised populations, i.e. the elderly, pregnant women,
young children, organ transplant recipients, and HIV positive
individuals, are more susceptible to infection by food borne
pathogens than healthy young and middle-aged adults
(Loaharanu, 1996).
The intake of fruit and vegetable juices are
recommended for a healthy diet and various health effects
(Williams, 1995). Healthy dietary behaviors such as an
increased consumption of fruit; vegetables and 100% fruit and
vegetable juice and a reduced consumption of sweetened
beverages have been related to a lower chronic disease risk and
body weight (Ludwig et al ., 2001).
A number of studies have supported an association
between a high consumption of vegetables and fruits and low
Introduction
incidences of certain chronic diseases (Lir et al., 2000).

Lifestyle-related diseases such as cancer, heart disease, and
hypertension have increased recently. These diseases are closely
related to the change of dietary habits; increase of animal
protein and fat intakes and a decrease of the intake of dietary
fibers. As a result, various original raw fruit and vegetable
juices, as a functional food, have gained popularity to reduce the
incidence of these diseases. The consumption of raw (
unpasteurized ) fruit and vegetable juices has increased in recent
years due to their characteristics of freshness, high vitamins
content, low calorie contribution and an active promotion of
fruits and vegetables and their derivatives as important
component of healthy diet ( Harris et al., 2003 ).
Fresh fruits and vegetables juices are highly
contaminated by microorganisms rendering its shelf-life very
short even at refrigeration temperature. The microbial flora in
the vegetable juice is thus likely to be similar to the flora in raw
fruits and vegetables. Many microorganisms, in particular acidloving or acid-tolerant bacteria and fungi (yeasts and molds),
can use fruit as substrate and cause spoilage, producing off
flavors and odors and discoloration of the product. The
unpasteurized fresh juices can support the growth of both
spoilage and pathogenic bacteria because they have high water
activity (aw) and enough nutrients for microbial growth. If a
bacterial pathogen is present on or in the product or on any
processing surface, it will have an almost unlimited food source
for growth in the unpasteurized juice, resulting in a foodborne
outbreak (Cook et al., 1998). Therefore, raw fruit and vegetable
juices should undergo some type of processing to inactivate
most of the microorganisms. The main requirement that a
process must meet is to ensure the microbial safety of the
product while preserving the sensory and nutritional
characteristics to obtain products similar to fresh fruit and
vegetable juices.
Introduction
The conventional methods usually used for reducing

microbiological contamination level of fresh fruit and vegetable
juices are washing practice of raw materials in water with a
combination of ozone and/or chlorine treatment. However, it has
a limited effect on microflora and sometimes can contaminate
the products. Nguyen-the and Carlin (1994) reported that
chlorine cannot be relied to eliminate pathogenic
microorganisms such as Listeria monocytogenes. On the other
hand, the use of heat can destroy nutrients such as thermally
labile vitamins and also the components responsible for a
product flavor and taste. Therefore, a non-thermal pasteurization
technology such high hydrostatic pressure (HHP) is required to
apply in the processing of fresh vegetable juice to avoid the
deleterious effects that heat has on the flavor, color, and nutrient
value of the foods (Barbosa-Canovas et al., 1998). However,
the HHP method still has several problems such as the
limitations of a mass production and an increased cost. For the
reasons, an industrial application of HPP is still limited. Ionizing
radiation, another non-thermal sterilization method, has
approved as highly effective in inactivating food microorganism
and has many advantages for industrial use. Recently, it has
offered a safe alternative as a decontamination method of food
and public health products (Niemira et al., 2001 and Hammad
et al., 2010).
The prescribed advantages of ionizing radiation (gamma
radiation, electron beam, x-rays ) arises from its ability to
destroy viable cells of microorganisms without arising the
temperature of the product, hence keeping its freshness,
nutrients, chemical and physical properties.
Food irradiation is a means of food preservation that has
been in development since the early part of the 20th century. If
applied properly, irradiation can be an effective way to reduce
the incidence of foodborne diseases and also inactivates food
Introduction
spoilage organisms, including bacteria, molds, and yeasts in our

food supply (Morehouse, 2002). At the same time, it can widen
the variety of available meals for immunocompromised patients,
by allowing the inclusion of some products normally considered
as high risk due to their microbial load, but that can be
nutritionally or psychologically adequate (Adeil Pietranera et
al., 2003).
The present study was designed to use an alternative
process, to heat pasteurization, that would reduce the microbial
load and eliminate pathogenic microorganisms from raw fresh
juices with maintenance of its freshness, sensory attributes and
most important nutrition. The irradiation treatment employed for
this study should not necessarily render products completely
sterile, but result in low-microbe and pathogen free meals
depending on the condition of patients. Thus, the main
objectives of the present study are:
1) Using -irradiation to improve the microbiological quality,
eliminate foodborne pathogens, and extend the shelf life of
some fresh fruit and vegetable juices (pomegranate, tomato
and carrot juices) to be used for immunocompromised
patient.
2) Investigate the microbiological, nutritional, physiochemical
and sensory quality of juices treated by gamma irradiation
during storage at refrigeration temperature.
3) Using cinnamaldehyde in combination with -radiation to
extend the shelf-life of carrot juice at refrigeration
temperature.
Introduction
Review
of Literature
Review of Literature
1. Immunocompromised patients
The immunocompromised host is a person with one or
more defects in the body's normal defense mechanisms because
of an immunodeficiency disorder or other disease or as the
result of the administration of immunosuppressive drugs or
radiation, that predispose him to infections, often lifethreatening, which would not otherwise occur. Patients
undergoing cancer therapy, transplant procedures and human
immunodeficiency virus (HIV) infection are more
immunocompromised than the aged, infant, pregnant or those
with multiple medical problems ( Butterweck, 1995).
Patients undergoing allogeneic transplant receive high
doses of chemotherapy with or without the addition of
radiotherapy and prolonged periods on anti-rejection drugs;
therefore, immunosuppression is generally lengthy and severe.
Patients undergoing autologous transplant receive chemotherapy
resulting in a shorter of severe neutropenia and less overall
immunosuppression.{Neutropenia is a consequence of hematological malignancies and the chemotherapy regimens used to
treat them} (Bodey et al., 1966). Finberg and Talcott (1999)
stated that treatment modalities such as stem cell transplant
(SCT), which can provide long term remission or cure for many
patients, often results in periods of profound neutropenia.
Neutropenia is a particularly major risk factor for the
development and outcome of fungal infections. Patients with
cancer and/or hemopoietic cell transplantation usually develop
neutropenia secondary to their malignancies or to their
chemotherapy treatment. Other risk factors are corticosteroid
Introduction
Review
of Literature
treatment and phagocytic dysfunction occurring in

immuno- suppressing conditions, including bone marrow
transplantation (BMT) and infection with HIV (Roilides et al.,
2002).
2. Immunocompromised patients problems
One major side effect of chemotherapy is the
development of infection as a result of neutropenia, or lowering
of the white blood cell count that result from damage to the
bone marrow and severe marrow suppression (DeMille et al.,
2006). When the absolute neutrophil count (ANC) is less than
0.5x109/l, there is a significant rise in infection risk with the risk
increasing in proportion to the length and severity of
neutopenia. Neutropenic patients with cancer constitute a
heterogeneous population with a variable risk for developing
serious complications (Paesmans, 2000).
Immunocompromised patients are at high risk for
opportunistic infections. Traditionally, there are infections that
arise from endogenous reactivation of latent infections, and
nosocomial transmission or nosocomial infections. Nosocomial
infections are infections that develop within a hospital and
produced by microorganism acquired hospitalization. There are
many sources of pathogens including decorative plants, hospital
workers, medical devices, food and water.
Children with HIV infection are at risk of developing a
wide spectrum of pathogens (Chanock and Pizzo, 1995).
Epidemiological investigations and air sampling experiments
lend support to the hypothesis that reinfection with a pathogenic
organism does occur, airborne transmission is possible and
nosocomial spread is a plausible explanation for new cases in
immunocompromised population (Lundgren et al., 1997).
These observations support the view that infectious
complications in
Introduction
Review
of Literature
immunocompromised patients are exogenous in origin. The fact

that infectious complications in immunocompromised patients
are often predictable and may be preventable makes infection
control practices a very important step in the improvement of
the quality of care provided to such patients (Risi and
Tomascak, 1998). Daw and Falkiner (1994) reported that
Gram - negative bacilli are a frequent cause of death associated
with infection and generally arise from the patients own
endogenous flora or are acquired during hospitalization.
Maguire et al. (2000) noticed that outbreaks of
foodborne infection in hospitals are associated with high attack
rates and disruption of services. Moreover, for
immunocompromised patients, the potential for food to cause
infection is even greater and hospitals may impose dietary
restrictions to limit pathogen exposure.
Adeil Pietraneraa et al. (2003) found that immunosuppressed patients are very likely to acquire microbial
foodborne diseases, since their natural defences are below what
is considered as normal limits. This makes their food intake
very restricted, avoiding all those products that could be a
source of microorganisms. Therefore patients with neutropenia
often are told to avoid fresh fruits and vegetables (Fishman and
Mrozek-Orlowski, 1999). Bibbington et al. (1993) reported
that the restriction of fruits, vegetables and salads reduces the
intake of water soluble vitamins and patients may find that diets
restricted to frozen meals or tinned foods are unpalatable and
unfamiliar. Therefore, the food service should be flexible and
designed to provide a variety of foods.
Introduction
Review
of Literature
3. Immunocompromised patients recommended diets

Imposing dietary restrictions for immunocompromised
patients continues to be widely used despite the lack of any
rigorous research support it (Wilson, 2002). These restrictions
are referred to throughout the literature and different institutions
by a variety of titles such as low bacterial diet, low microbial
diet, clean diet or neutropenic diet. The low bacterial diet was
designed to reduce pathogens by excluding foods that may cause
systemic bacteraemia or Gram-negative sepsis via the hosts
gastrointestinal tract.
Pizzo et al. (1982) evaluated the microbiologic content
of 236 commercially available food items. Since then, the low
bacterial diet has changed from a sterile diet prepared under
laminar-flow hoods to a more liberal diet that avoids high-risk
foods and emphasizes safety in food handling practices.
Problems of limited availability of single-serve sterile foods,
lack of standardization of recipes, and low patient acceptance of
autoclaved sterile foods were reported as reasons for the move
toward less stringent dietary procedures (Dezenhall et al.,
1987).
In order to reduce the risk of infection, several
preventive measures have been adopted (Schwartz and Perry,
1966). Fundamentally; all of these measures were designed to
prevent either acquisition of gram negative bacilli or fungal
pathogens from the environment or the translocation of these
potential pathogens across the mucosal barrier of the gut. These
measures include protective (or reverse) isolation, antibiotic
prophylaxis with antibiotics which selectively eradicate the
aerobic gram-negative bacilli and yeasts from the gut flora and
finally the use of low-bacterial diets (LBDs). Several studies
have documented the frequent isolation of Enterobacteriaceae
from food (Hamilton-Miller and Shah, 2001).
Introduction
Review
of Literature
Lowmicro-organism diets are indicated to prevent

sepsis in transplant recipients. These diets should comply with
the following basic guidelines:
* Omits raw vegetables, salads, soft cheeses, raw meat products,
most fresh fruits, tap water and spices (Van tiel et al., 2007).
* Avoid foods containing yeasts and gram-negative bacteria.
* Practice safe food handling and preparation techniques to
avoid contamination.
* Avoid foods intrinsically contaminated with microorganisms,
such as raw eggs, raw or rare-cooked meat, fish, seafood, and
unpasteurized milk (Martin-Salces et al., 2008).
Dietitians who work with leukaemic patients are often
asked by medical and nursing staff to advice on low microbial
or sterile diets for patients undergoing bone marrow
transplants (BMT). Sterile diets were also prescribed and are
defined as food free from bacterial or fungal growth after 7 days
incubation (Frankmann et al., 1984). These diets were
achieved by autoclaving or irradiating foods, necessitating
specialist, labour-intensive facilities.
Severly immunocompromised patients such as in cancer
treatment and organ transplants need sterile diets. Other patients
that are immunocompromised do not require sterile diets. These
patients may require a diet that is pathogens free and are also
low-microbial diets(Butterweck, 1995). However, many
investigators recommended clean-diet or low-microbial diets for
immuno-compromised patients instead of sterile diets (Aker
and Cheney, 1983; Somerville, 1986).
Introduction
Review
of Literature
4. Importance of fresh fruits and vegetables and their juices

for human health
The association between fresh fruit and vegetable
consumption and cancer risk has been reported in hundreds of
epidemiological studies (IARC, 2003).
It has been
hypothesized that the wide variety of nutrients and bioactive
compounds in fresh fruits and vegetables can inhibit
carcinogenesis during the initiation, promotion, and progression
stages. The nutrients and bioactive compounds present in fruits
and vegetables have been shown to inhibit the deactivation of
pro-carcinogens, induce detoxification pathways, affect the cell
cycle by regulating cell cycle progression, influence cell-to-cell
communication, quench free radicals, stimulate the immune
system, modulate hormone metabolism, and dilute and bind
carcinogens, all of which should help to prevent the
development of cancer by promoting the proliferation of
lymphocytes, stimulating macrophage phago-cytosis, and
enhancing the activity of natural killer cells, these nutrients may
protect cells against microbes, bacteria, and viral and fungal
agents (Head, 1998).
Fruits and vegetables are also rich sources of
antioxidants (Wang et al., 1996), which help to quench
exogenous and endogenous free radicals. If the free radicals
remain in an oxidative state, they may cause oxidative damage
to nucleotides, proteins, and cell membranes and result in the
initiation of carcinogenesis. Antioxidants are a diverse group of
compounds that act against oxidative damage induced in the
body. These antioxidants are not present in abundance in the
normal diet, but have to be increased mainly by plant sources.
Reports available have shown that antioxidants play a
significant role in the prevention of cancer, cardiovascular
disease, cataract formation, the aging process, inflammatory
diseases, and a wide range of
10
Introduction
Review
of Literature
neurological disorders. Polidori (2003) indicated that foods

containing high levels of antioxidants may also slow the
progression of Alzheimers disease (AD), possibly by
preventing or neutralizing the damaging effects of free radicals.
Fresh and processed fruits and food products also
contain high levels of a diverse range of phytochemicals of
which polyphenols including hydrolyzable tannins [ellagitannins
(ETs)
and
gallotannins]
and
condensed
tannins
(proanthocyanidins), and anthocyanins and other flavonoids
make up a large proportion ( Gu et al., 2004 ). According to
Liu (2004), it should be noted that phytochemical extracts from
fruits and vegetables have strong antioxidant and
antiproliferative activities, and the foremost part of total
antioxidant activity is from the combination of these
phytochemicals. This author also proposed that the additive and
synergistic effects of phytochemicals in fruits and vegetables are
responsible for potent antioxidant and anticancer activities, and
the benefit of a diet rich in fruits and vegetables is attributed to
the complex mixture of phytochemicals present in whole foods.
Phytochemicals can be present in abundance in
vegetables (e.g., tomatoes, green leafy vegetables, and broccoli),
fruits (e.g., berries, olives, oranges, mangosteens, and grapes),
nuts (e.g., hazelnuts and almonds), herbs (e.g., sweet clover,
ginseng, sage, and rosemary), and flowers (e.g., hibiscus and
marigold), as well as in medicinal plants (e.g., periwinkle and
opium). However, depending on their composition and
presence, different phytochemicals are found in different plant
species and at different concentrations. For example, the
phytochemicals present in the forms of alkaloids (e.g., caffeine
and threbromine), carotenoids (e.g., lycophene), coumestans
(e.g., flavon-3-ols), isoflavones (e.g., genistein), phenolic acids
(e.g., capsaicin, gallic
11
Introduction
Review
of Literature
acid, and tannic acid), etc., might vary in concentration among

different plant species, and may either be present or totally
absent. Also, for example, fruits like orange and strawberry
have rich contents of flavonoids like hesperidin and
anthocyanins (Wimsen et al., 2005), while vegetables mainly
contain apigentin, quercetin, quinones, luteolin, etc. (Miean and
Mohamed, 2001). Fresh fruits and vegetables are widely
consumed fresh of squeezed into juices.
WCRF (2007) emphasized that diets rich in fruits and
vegetables are protective against disease and populations that
consume such diets have higher plasma antioxidant status and
exhibit lower risk of cancer and cardiovascular disease. SauraCalixto and Goi (2009) reported that dietary fiber may also
include associated bioactive compounds such as polyphenols
and carotenoids. Bioactive compounds associated with dietary
fiber may explain the qualitative nutritional differences found in
a healthy dietary pattern, such as the diet in Mediterranean
countries and other fruits and vegetables are considered rich
sources of some essential dietary micronutrients and dietary
fiber, and more recently they have been recognized as important
sources of a wide array of phytochemicals that individually or in
combination may benefit health (Yahia, 2010).
Wang et al. (2011) reported that antioxidant properties
of some bioactive components in plant-based foods have been
proposed to be capable of controlling the disturbances of the
normal redox state within the human body result from oxidative
stress and that antioxidants found in three categories of plantbased foods (fruits, vegetables and legume) and mechanisms
that these antioxidants may use in promoting cardio-health.
Since different food categories possess different bioactive
compounds with various antioxidant capacities, specific foods,
when
12
Introduction
Review
of Literature
consumed together, may produce synergistic antioxidant

interactions and in turn have more positive physiological effects
on cardio-health than when consumed alone.
4.1. Pomegranate juice
Pomegranate (Punica granatum L.) fruits are widely
consumed fresh and in beverage forms (Gil et al., 2000). The
medicinal properties of pomegranate are described by all major
religions and by folk medicine (Langley, 2000). Pomegranate
fruit extract is a rich source of 2 types of poly phenolic
compounds: anthocyanins (derived from delphinidin, cyanidin
and pelargonidin), which give red color to the fruit and juice,
and hydrolysable tannins (i.e. punicalin, pedunculagin,
punicalagin, gallagic and ellagicacid esters of glucose), which
account for 92% of the antioxidant activity of the whole fruit.
These compounds are known for their properties in scavenging
free radicals and inhibiting lipid oxidation in vitro (Gil et al.,
2000). In addition to its antioxidant activity, it has antimicrobial,
antibacterial, antiviral, antifungal and antimutagenic properties
as well as beneficial effects on the oral and cardiovascular
diseases (Cook and Samman, 1996).
Pomegranate juice is a rich source of different phenolic
compounds, with anthocyanins to be one of the most important
classes (Du et al., 1975). The antioxidant level in pomegranate
juice was found to be higher than that in other natural juices
such as blueberry, cranberry, and orange (Aviram et al., 2000).
Minerals in the juice and seed include Fe, relatively prevalent,
but not in so high concentrations as in watermelon, and Ca, Ce,
Cl, Co, Cr, Cs, Cu, K, Mg, Mn, Mo, Na, Rb, Sc, Se, Sn, Sr, and
Zn (Waheed et al., 2004).
13
Introduction
Review
of Literature
4.2. Tomato juice

A large part of the world's tomato crop (Solanum
lycopersicum) is processed into tomato juice and other products.
Epidemiological studies have shown that increased consumption
of tomato and tomato juices may reduce the risk of certain types
of cancer, such as prostate, lung and stomach cancer
(Giovannucci, 1999). Tomato ranked first as a source of
lycopene (71.6%), second source as a source of vitamin C
(12.0%), provitamin A carotenoides ( 14.6%) and -carotene
(17.2%) and third as a source of vitamin E (6.0%) as reported by
Garcia-Closas et al. (2004). Lycopene, the pigment principally
responsible for the characteristic deep-red color of ripe tomato
fruits and tomato products, has received much attention in
recent years because of its beneficial effect in the treatment of
diseases (Shi and Le Maguer, 2000). Inparticular, tomatoes
represent the most important source of lycopene, a carotenoid
with a high oxygen-radical scavenging and quenching capacity
(Dumas et al., 2003). The positive role of tomato on human
health has been ascribed principally to its vitamin C content and
carotenoids constituents, particularly lycopene and -carotene,
that accumulate in plasma and tissues in relation to tomato
(Dorais et
al., 2008).
Fig ( Fig. (1): Carotenoid profile of tomato products (RoldanGutierrez and Luque de castro, 2007).
14
Introduction
Review
of Literature
4.3. Carrot juice

Carrot juices as natural extract from carrot crop (Daucus
Carota) enjoys high consumer acceptance on account of its taste
and nutritional benefits. Carrot juice is however preferred as
fresh-squeezed. Although carrots are assumed to be safe, the
consumption of unpasteurized juices may be dangerous since
they could be a potential source of bacterial pathogens (Ryu and
Beuchat, 1998). Outbreaks of food poisoning caused by
biological hazards have been traced back to the consumption of
carrots and carrot juice (Anon, 1999). Carrot juices are
preferably used as a natural source of provitamin A in the
production of alpha-tocopherol-beta carotene drinks (ATBCdrinks) because of its high content of -carotene (Marx et al.,
2000). Carrot is one of the most commonly used and wellknown vegetables in the everyday kitchen that is rich in
functional food components such as vitamins (A, D, B, E, C,
and K) and minerals (calcium, potassium, phosphorus, sodium,
and iron).
The carotenoids and other antioxidants present in carrot
play an important role in the inhibition and/or interruption of
oxidation processes, as well as in counterbalancing free radical
activities (Krinsky and Johnson, 2005). Therefore, carrot may
protect humans against certain types of cancer and
cardiovascular diseases.
4.4. Anthocyanin
Anthocyanins are plant pigments responsible for the
orange, red and blue colours of various fruits and vegetables
(Rentzsch et al., 2007).
Anthocyanins derived from
delphinidin, cyanidin and pelargonidin (Gil et al., 2000).
15
Introduction
Review
of Literature
Fig. (2): Structure of Anthocyanin

High anthocyanin content is appreciated due to its strong
chemopreventive activities such as antimutagenicity, antihypertension, antioxidative potential and reduction of liver injury (Du
et al., 1975). Anthocyanins, potent antioxidant flavonoids,
provide pomegranate juice with its brilliant color, which
increases in intensity during ripening (Hernandez et al., 1999),
and declines after pressing ( Miguel et al., 2004).
4.5. Lycopene
Lycopene is a vibrant red carotenoid that serves as an
intermediate for the biosynthesis of other carotenoids( Stahl
and Sies, 1996 ). It is an acyclic open-chain unsaturated
carotenoid found in moderate to high concentrations in such
foods as red tomatoes and processed tomato products,
watermelon, red grapefruit, and Brazilian guava, red pepper,
apricot, papaya, passion fruit, guava, carrots, pepper,
persimmon, balsam pear and a large number or redberries (e.g.,
cranberries) and fruits of Autumn olive.
16
Introduction
Review
of Literature
Fig.(3): Structure of all-trans Lycopene

Lycopene, a fat soluble carotenoid, is a precursor of carotene (Sandmann, 1994) and has at least twice the
antioxidant capacity of -carotene; it is an efficacious free
radical scavenger (DiMascio et al., 1989). Epidemiological
studies have indicated positive health benefits in consumption of
diets high in lycopene, and its presence in the diet positively
correlates with reduced cancer incidence (Gerster, 1997).
Although it has no provitamin A activity, lycopene is able to
function as an antioxidant and to quench singlet oxygen in vito.
The quenching constant of lycopene was found to be more than
double that of -carotene and 10 times more than that of tocopherol (Shi et al., 1999). In addition to its antioxidant
properties, lycopene has also been shown to induce cell to cell
communication ( Zhang et al., 1991) and to modulate hormonal
and immune systems and other metabolic pathways which may
also be responsible for the beneficial effects (Rao and Agarwal
, 1999). Extensive scientific studies have found that it appears to
have strong antioxidant capabilities that can inactivate free
radicals and reduce damage to the bodys cells, reducing the risk
from some human cancers and chronic diseases (Ghaffari and
Ghiasvand, 2006).
17
Introduction
Review
of Literature
Table (1): Shows the approximate content of lycopene in

different fruits and vegetables and other foods as
reported by Roldan-Gutierrez et al., 2007)
Food
Content ( mg / 100g )
Tomatoes, fresh
Tomatoes, cooked
Tomato sauce
Tomato paste
Tomato soup, condensed
Tomato powder
Tomato juice
Sun-dried tomato in oil
Pizza sauce, canned
Ketchup
Apricot
Apricot, canned
Apricot, dried
Grapefruit, raw pink
Guava, fresh
Guava, juice
Watermelon, fresh
Papaya, fresh
0.88-4.20
3.70
6.20
5.40-150.00
7.99
112.63-126.49
5.00-11.60
46.50
12.71
9.90-13.44
<0.01
0.06
0.86
3.36
5.40
3.34
2.30-7.20
2.00-5.30
4.6. Carotenoids
Valued for the pleasing yellow, orange or red color they
impart to many foods, carotenoids are also natural antioxidants
and as such contribute to the stability of foods. Moreover, aside
from the provitamin A activity, these compounds have healthpromoting effects: immuno-enhancement and reduction of the
risk of developing degenerative diseases such as cancer,
18
Introduction
Review
of Literature
cardiovascular diseases, cataract and macular degeneration

(Voutilainen et al., 2006). For a long time, quantification of
only -carotene or the major provitamin A carotenoids was
considered sufficient.
Fig. (4): Structure of carotene

Beta-carotene is a natural compound that is omnipresent
in yellow and green vegetables and fruits, such as carrot,
spinach, sweet potato, pumpkin, broccoli, peppers, pink
grapefruit, papaya and peach. Beta-carotene is also a coloring
agent that is ubiquitously added to food and drinks.
Furthermore, beta-carotene is an important source of vitamin A.
Beta-carotene is added to plant based margarines, to compensate
for their lack of vitamin A. More recently, a rice variety (golden
rice) has been made by genetic engineering that contains betacarotene to combat vitamin A deficiency in developing
countries. From the above it can be seen that beta-carotene is
widely present in the diet and that it may influence our health
(Keijer et al., 2005).
4.7. Ascorbic acid
Ascorbic acid is the most important vitamin for human
nutrition that is supplied by fruits and vegetables. Since
vegetables are an important source of vitamin C in the total diet,
it is important that any preservation method used to extend their
shelf life will not markedly affect the vitamin C content.
Ascorbic acid and the B vitamins can affect immune response.
19
Introduction
Review
of Literature
Prophylactic ascorbic acid has been used for upper

respiratory viral illness and for combating cold symptoms while
B vitamins may act as controlling factors in the rapidly dividing
effector cells of the immune system (Anonymous, 1990). Sato
and Miyata (2000) reported that vitamin C found in many fruits
and vegetables and organosulfur compounds found in onions,
leeks, and garlic may additionally stimulate the immune system.
Fig. (5): Structure of Ascorbic acid

5. Microbial contamination of food in hospitals
Outbreaks of food borne infection in hospitals are
associated with high attack rates and disruption of services
(Maguire et al., 2000). It was found that in 2002, hospitals in
the Netherlands were implicated in 9% of 281 gastroenteritis
outbreaks (Van Duynhoven et al., 2005). A foodborne
outbreak of Salmonella infection at a private hospital in London
in 1994 had an attack rate estimated to be 5% among the
approximately 200 patients and staff at risk (Maguire et al.,
2000). In Poland, the annual outbreaks of food poisoning and
foodborne infections in hospitals and sanatoria from 1985 to
1999 constituted from 1.5% to 6.3% of the total number of such
outbreaks in the country (Przybylska, 2001). Another study in a
national hospital in Costa
20
Introduction
Review
of Literature
Rica revealed that all tested salad samples were positive for
faecal coliforms (Jimenez et al., 2004). A study in a university
hospital in France showed that 10% of patients meals, all of
which were salads, had total viable bacteria counts above the
recommended limits as shown in table (2) ( Reglier-Poupet et
al., 2005 ). Also, poultry meat is an important vehicle for food
poisoning organisms, particularly Campylobacters and
Salmonellas (Adak et al., 2005).
Table (2): French recommended microbiological limits for different
itemsoffood(ReglierPoupetetal.,2005)
Bacteria number cfu/g
Acceptable
unsatisfactory
Good
Salads
Mesophilic organisms
Faecal coliforms
Meat
Faecal coliforms
Staphylococcus aureus
Anaerobes
Cooked plates and
plates with eggs
Faecal coliforms
Total coliforms
Anaerobes
Vegetables
Faecal coliforms
Total coliforms
Anaerobes
<105-1.5.106
<10-30
1.5.106-5.106
30-100
>5.106
>100
<5.105-1.5.106
<102-3.102
<102-3.102
<30-90
1.5.106-5.106
3.102-103
3.102-103
90-300
>5.106
>103
>103
>300
<3.105-9.105
<10-30
<103-3.103
<102-3.102
<30-90
9.105-3.106
30-100
3.103-104
3.102-103
90-300
>3.106
>100
>104
>103
>300
<5.105-1.5.106
<10-30
<103
<102-3.102
<30-90
1.5.106-5.106
30-100
3.103-104
3.102-103
90-300
>5.106
>100
>104
>103
>300
21
Introduction
Review
of Literature
One of the most important factors related to food borne

illnesses is the lack of knowledge on the part of food handlers or
consumers and negligence (despite knowledge) in safe food
handling (WHO, 2000). Aycicek et al. (2004) determined the
level of bacterial contamination on the hands of food handlers (n
=30), (n is the number of the food handlers), who work in the
kitchen of a military training hospital. A total of 180 samples
were collected from bare and gloved hands before and during
food preparation. A total of 16 different bacteria were isolated,
of which the most common was Staphylococcus aureus
(126/180; 70%), followed by coagulase-negative staphylococci
(102/180; 56.7%), Diphtheroid bacilli (39/180; 21.7%), Bacillus
spp. (19/180; 10.5%), and Escherichia coli (14/180; 7.8%).
Fifty-one of 60 (85%) gloved hand samples were collected
during work, 57 (95%) of the bare hand samples were collected
before work all of the bare hand samples collected during work
were positive. Poor hand hygiene was indicated by high levels
of S. aureus and E. coli on samples taken from bare and gloved
hands. Although bacterial loads on gloved hand samples were
found to be significantly lower (p < 0:05) than ungloved hand
samples, these loads were not within acceptable limits. These
results show that the hands of food handlers are an important
contamination source in this establishment.
Njobeh et al. (2009) investigated that a total of 95
human food samples to evaluate the incidence of fungal
contamination in Cameroon. The investigation revealed the
predominance of Aspergillus and Penicillium with 96% of
samples contaminated with at least one species of these fungi,
whereas the incidence of co-contamination of samples was 85%.
Aspergillus flavus and Aspergillus parasiticus were the most
predominant species contaminating mainly maize and peanuts.
In addition, Penicillium crustosum and Penicillium polonicum
were the most common
22
Introduction
Review
of Literature
contaminants belonging to the genus Penicillium. On the other

hand, Aspergillus ochraceus registered a low incidence rate of
5%, including other members of the Aspergillus group. Other
members of the genera Rhizopus and Alternaria spp. were also
registered in the study. A majority of fungal strains of A.
ochraceus, A. parasiticus, P. crustosum and P. polonicum
isolated were toxigenic, producing the mycotoxins tested for,
while none was detected in cultures of A. fumigates.
Malheiros
et al. ( 2010) stated that Crosscontamination through working surfaces is widely reported,
specially the cases of transmission of enteric pathogens and
other microbial hazards such as Listeria monocytogenes or
Staphylococcus aureus to ready-to-eat ( RTE ) foods.
Rodriguez et al. (2011 ) evaluated that lettuce salads samples
presented high counts of MAB (mesophilic aerobic bacteria )
and total coliforms (104 to 105 and 10 to 104 cfu/g, respectively),
Neither Listeria spp. nor Salmonella spp. were detected in any
food sample of chilled ready-to-eat (RTE) foods in hospital
foodservices.
Fresh fruit and vegetables are essential components of
the human diet and there is considerable evidence of the health
and nutritional benefits associated with the consumption of fresh
fruit and vegetables. Beuchat ( 1996 ) discussed the incidence
of human pathogens on fresh produce, citing contact with
contaminated soil, water, compost, harvesting or processing
equipment, or human handlers as mean by which produce may
become contaminated. Sizer and Balasubramaniam (1999)
indicated that the microbial populations on and in fruits and
vegetables were carried into derivative juices or fresh cut
products during processing. Plant parts such as leaves, stems,
23
Introduction
Review
of Literature
fruits and roots typically support 103 to 106 colony forming units
(CFUs) per gram of plant tissue (Mercier and Lindow, 2000).
Fresh produce can be a vehicle for the transmission of
bacterial, parasitic and viral pathogens capable of causing
human illness and a number of reports refer to raw vegetables
harbouring potential foodborne pathogens (Nguyen-the and
Carlin, 1994; Beuchat, 1996).
Listeria monocytogenes
(Schlech et al., 1983), Salmonella (Doyle, 1990), and
Escherichia coli (Nguyen-the and Carlin, 1994), are most
frequently associated with fresh produce fruits and vegetables.
There are a number of reports indicating that raw vegetables
may harbor the potential of food-borne pathogens (Beuchat,
1996). Also, the pathogens most frequently linked to producerelated outbreaks include bacteria (Salmonella, Escherichia
coli), viruses (hepatitis A), and parasites (Cryptosporidium,
Cyclospora) (Tauxe et al., 1997).
Beuchat (1998) indicated that vegetables can become
contaminated with microorganisms capable of causing human
diseases while still in the fields or during harvesting or postharvest handling in food services establishments. Todd (1999)
postulated that fresh fruits and vegetables appear to carry
several species of Gram-negative rods as part of their natural
flora. It has been demonstrated that several kinds of salads were
colonized by Pseudomonas aeuroginosa, E. coli and Klebsiella
spp. Olsen et al. (2000) demonstrated that Salmonella and E.
coli O157:H7 is being the leading causes of produce-related
outbreaks in the USA.
Lugauskas et al. (2005) investigated toxin producing
fungi in fresh fruits sampled in Lithuania. Penicillium
expansum, Sclerotinia sclerotiorum, Alternaria alternata, and
Penicillium italicum were the prevalent fungal species, while
Penicillium expansum, Aspergillus niger, Rhizopus oryzae and
Penicillium italicum dominated on dried fruits.
24
Introduction
Review
of Literature
While, Mukherjee et al. (2006) postulated that the

incidence of foodborne outbreaks caused by contaminated fresh
fruit and vegetables has increased in recent years. Abu-El Nour
(2007) found that fresh-cut carrot samples had high level of
microbial counts (1.4x104 - 2.4x106 cfu/g) including lactic acid
bacteria, coliform bacteria, Escherichia coli and Staphylococcus
aureus. Badosa et al. (2008) emphasized that bacterial
pathogens usually found in contaminated vegetable and fruit
products include Salmonella, Escherichia coli O157:H7,
Listeria monocytogenes, and Staphylococcus aureus. Hell et al.
( 2009 ) stated that fungal contamination was evaluated after
plating on selective media with a total of 561 fungal isolates
identified, ranging from 18 in tomato and 218 in baobab leaves.
Baobab leaves, followed by hot chilli and okra showed high
incidence of fungal contamination compared to the other dried
vegetables, while shelled melon seeds, onion leaves and dried
tomato had lower levels of fungal contamination. Ponniah et al.
(2010) found that Listeria spp. and Listeria monocytogenes
could be detected in 33.3% and 22.5% of the vegetables,
respectively. Verhoeff-Bakkenes et al. (2011) reported that
thirteen of the 5640 vegetable and fruit samples were
Campylobacter positive.
juices
In the recent years, the tendency of consumers to fresh
fruit and vegetable juices has been increased due to their better
organoleptic properties and high vitamin content than
pasteurized ones. However, these products without a minimal
processing may be potential source of microbiological diseases,
since the low acidity (pH 5.26.7) and high water activity (0.97
0.99) of these juices can favor the growth of pathogenic
microorganisms (USFDA, 2001).
25
Introduction
Review
of Literature
The unpasteurized fresh fruit and vegetable juices can

support the growth of a variety of bacterial pathogens coming
from damaged raw fruits and vegetables and through crosscontamination during processing. Wells and Butterfield (1997)
showed that pathogenic microorganisms such as Salmonella
could grow in the wound area of a damaged, intact fruits or
vegetables. If bacterial pathogens are present on or in the
produce or any processing surface, it will have an almost
unlimited food source for growth in the unpasteurized juice,
resulting in a food borne outbreak (Cook et al., 1998).
Fruit juices contain various concentrations of sucrose,
which constitutes a very important component of the medium
for the growth of both spoilage and pathogenic microorganisms
especially fungi. Microbial spoilage is a serious problem for the
food industry as fungal contamination can occur during
processing as well as handling of the end products. Since yeasts
can generally resist extreme conditions better than bacteria, they
are often found in products with low pH and in those containing
preservatives. Especially yeast spoilage has increased in recent
years as a result of lower doses of preservatives and milder
preservation processes, required for higher standards of food
quality (Koc et al., 2007).
Grinbaum et al. (1994) reported that yeast spoilage of
fruit juice can result in formation of haze, production of CO2
and off-odors, and changes in color. Goto et al. (2003) reported
the spoilage of fruit juices/beverages has been associated with
four
species
of
molds:
Alicyclobacillus
acidoterrestris,Alicyclobacillus pomorum, Alicyclobacillus
herbarius and A. Acidiphillus. Guerrero-Beltrn and BarbosaCnovas (2005) found that yeasts, molds, lactobacillus,
leuconostoc and thermophilic Bacillus are common spoilage
microorganisms of orange juice,
26
Introduction
Review
of Literature
Escherichia coli, Saccharomyces cerevisiae and Listeria

innocua are associated with apple juice or apple cider spoilage
Tournas et al. (2006) found that microorganisms, in particular
acid tolerant bacteria and fungi such as yeasts and molds, can
easily spoil fruit juices. Song et al. (2007) found that the
contaminating bacteria in the juices ranged from 106 to 107
CFU/ml in carrot and kale juice. Meanwhile, Alighourchi et al.
( 2008 ) found that the initial mean populations of the total
bacterial and fungi counts for fresh juices of two pomegranate
varieties were 4.7x103 , 3.4x103 and 2.0x103, 1.0x103 cfu/ml,
respectively. Hsu et al. (2008) reported that the tomato juice
proved to be naturally contaminated by the microorganisms
normally occurring in tomato juice, at the same levels as those
found in the tomato product, total viable count, Enterobacteria,
lactic acid bacteria and mold and yeasts were 4.1, 2.1, 4.2 and
3.7 Log cfu/ml, respectively. On a worldwide basis, the
incidence of Alicyclobacillus spp. has been reported mainly in
apple and orange juices and pear concentrate (Groenewald et
al., 2009).
8. Methods used to reduce the hazards of foodborne
infection for immunocompromised patients
Methods such as heating, cooling, freezing, drying,
freeze-drying,
irradiation,
high
hydrostatic
pressure,
fermentation, or the addition of antimicrobials and chemicals are
commonly used to control microbial contamination and
pathogens of patient diet. After these treatments, one population
of microorganisms may be killed, another population may
survive (non-injured), and a third population may be sublethally injured (Wu et al., 2001).
8.1. Thermal treatments
Heating (boiling, grilling, baking, and frying) is applied
to food to enhance its flavour and taste, inactivate pathogenic
27
Introduction
Review
of Literature
microorganisms and increase shelf life (Bognar, 1998). On the

other hand, the use of the microwave oven for cooking has
increased greatly during recent decades (Arias et al., 2003). But
it was found that cooking brings about a number of changes in
physical characteristics and chemical composition of vegetables
and other food materials. Cooking processes also produce some
structural changes in dietary fiber components of various
vegetables (Sukhwant et al., 1992).
Ben (1999) indicated that heat process can promote
changes in concentration of volatiles or trace elements. Such
losses of various macro or micro-nutrients and trace elements
may be detrimental for the human health. Zhang and Hamauzu
(2004) considered that heat preservation is one of the oldest
forms of preservation known to man and has the potential to
provide barriers to reduce microorganisms and inhibit enzyme
activity, but this treatment is incompatible with fresh processed
plant food since heat is associated with destruction of flavour,
texture, color, nutritional quality and antioxidant activity.
Pasteurization is a heat treatment which kills pan of the
vegetative microorganisms present in the food and relies heavily
on handling and storage conditions to further minimize bacterial
growth (Karel et al., 1975). It is the treatment of packaged
foods to temperatures below 100 C over a given time to
eliminate pathogenic microorganisms which may grow under
certain storage conditions. Pasteurization has been used to
prevent foodborne diseases associated with dairy products, milk,
liquid egg, fruit juices (Lalaguna, 2003). During the processing,
however, a potential hazard may occur that can be caused by
addition of contaminated ingredients or improper handling of
the final products, including an abuse of the storage temperature
(Jo et al., 2007).
28
Introduction
Review
of Literature
8.2. Refrigeration
With increasing world population and the need to supply
people with fresh and healthy food, food preservation becomes
increasingly necessary in order to increase the shelf life and
maintain the nutritional value, texture and flavour of food. A
main challenge in this respect is to maintain a stable and
sufficiently low temperature which is often more difficult in
fresh foods than in frozen foods. Studies on the chill chain have
shown that it is a challenge to maintain an acceptable
temperature during distribution and storage of food products
(Aune, 2003).
Controlling microbial activities is the key to extension of
shelf-life during processing, distribution and storage of food.
Shelf life is the time during which the product will remain safe,
to retain the sensory, chemical, physical and microbiological
characteristics, and comply with any label declaration of
nutritional data (Kilcast and Subramaniam, 2000).
Temperature is one of the most important parameters affecting
the growth of microorganisms and shelf-life extensions of
(Brand et al., 1999). The most methods used for shelf-life
extension of foods include refrigerated ice storage between 0 C
and 4 C, superchilled storage in the range of -1 to -4 C, by
means of slurry ice or in superchilled chambers without ice, and
frozen storage at -18 to -40 C (Gallart-Jornet et al., 2007).
Paull (1999) reported that low temperature (0 C and 4 C) has
been used to extend the shelf life of temperate fruits and
vegetables and their juices but the shelf-life was limited. Song et
al. (2007) found that the shelf life of natural vegetable juices
rarely exceed 1 day even under a cold chain system.
Refrigeration throughout the production chain up to
consumption is a fundamental importance in extending the shelf
life of fresh cutproducts.The microbiological and safety quality
of extended shelf life
29
Introduction
Review
of Literature
refrigerated foods continues to be an issue for processors and

the food preparer (IFT, 1998).
8.3. Chemical additives
The use of chemical additives in food and liquid has
increased immensely in the last decades. The main reason, given
by food manufacturers for using additives is that food would
spoil in a short time without additives (Shibamoto et al., 1993).
However, there is research asking to ban the usage of all
unnecessary additives consumed by infants and young children
(Tuormaa, 1994). Also, to prevent the lipid peroxidation in fats
and oils, synthetic antioxidants have been used as food additives
for over 50 years ( Cuvielier et al., 1994 ). Food additives have
been implicated as aetiological factors in many different disease
states. Concern arose from a suggested link with food additives
and hyperactivity in children. They have also been implicated in
many other disease states (Young, 1997). Some countries
already limited some food products in schools because
ingredients or additives of those products may cause
health/behavior problems in children (Fried and Nestle, 2002).
The additives have many disadvantages and risk to the
health and the life of the individual for example evidence has
been presented that ascorbic acid can interact with benzoic acid
in the presence of a transition- metal catalyst to yield benzene, a
known carcinogen (Gardner and Lawrence, 1993).
8.4. High hydrostatic pressure processing (HHP)
High hydrostatic pressure processing (HHP) or High
pressure processing (HPP) is a non thermal food preservation
technique for microbial and enzyme inactivation with reduced
effects on nutritional and quality parameters when compared to
thermal treatments (Tiwari et al., 2009). HHP has been
proposed
30
Introduction
Review
of Literature
for application to milk, vegetable juices and other food

(Considine et al., 2008). However, HHP method still has
several problems such as limitation of the mass production and
an increase cost. Rendueles et al. (2011) reported that the
introduction of an HHP step in the food manufacturing process
requires a careful assessment of microbiological risks. The
process safety assurance is enhanced when adequate hazard
identification is performed. As with other treatments, there is
also a need to have tools and mechanisms in place to monitor,
optimize and validate the process, and procedures to assess and
model the lethal effect of the treatment.
8.5. Ionizing Radiation
Decontamination of food by ionizing radiation is a safe,
efficient, environmentally clean and energy efficient process.
Irradiation is used to inactivate food-borne microorganisms, to
reduce quality losses during storage and to guarantee the
hygienic quality of several foodstuffs such as poultry, meat,
spices, herbs, sea foods, vegetables and fruits etc. ( Moy
and Wong, 1996). In recent decades, food irradiation has
become one of the most discussed technologies for food safety
and shelf-life. The purpose of food irradiation is the same as for
freezing, high temperature treatment and chemical treatment, i.e.
removal of spoilage and pathogenic microorganisms causing
food spoilage and human diseases. The aim is to prolong the
shelf-life of foods kept under various conditions such as in
shops and households, and to eliminate pathogenic organisms
that cause disease as a result of food consumption (Lee et al.,
2004).
31
Introduction
Review
of Literature
9. Treatment of food by irradiation

Irradiation has been recommended as a method for
preparing foods for hospital patients requiring sterile diets or
low microbial diet as a result of intensive therapy or disease that
has resulted in suppression of the immune system. It has a
number of advantages over other methods and in recognition of
this, in the United Kingdom; the use of irradiated foods for
hospital patients has been specifically exempted from regulatory
control. Due to a number of factors there is a move away from
keeping patients in a sterile environment; however, irradiation
may still have a role to play for vulnerable and high-risk patients
(Pryke and Taylor, 1995). Immunosuppressed patients are very
likely to acquire microbial food borne diseases, since their
natural defences are below what is considered as normal
limits. This makes their food intake very restricted, avoiding
all those products that could be a source of microorganisms.
Ionizing radiation applied at sub-sterilizing doses represents a
good choice in order to achieve clean diets for
immunosuppressed patients and at the same time, it can widen
the variety of available meals for these patients, allowing the
inclusion of some products normally considered as high risk
due to their microbial load, but that can be nutritionally or
psychologically adequate.
Several researchers have proposed that ionizing
irradiation was a suitable method to control the microorganisms
of fruits, fresh fruit juices, fresh-cut vegetables, salads, sprouts,
seeds and other minimally processed foods (Buchanan et al.,
1998) as a non-thermal sterilization method. Beuchat (1998)
stated that ionizing radiation could be an extremely effective
tool in reducing the populations of pathogenic microorganisms
and parasites from raw fruits and vegetables. In 1997, a Joint
FAO/IAEA/WHO Study Group was convened to assess the
safety
32
Introduction
Review
of Literature
and nutritional adequacy of food irradiated to doses above 10

kGy. They concluded that food irradiated to any dose
appropriate to achieve the intended technological objective is
both safe to consume and nutritionally adequate (WHO, 1999).
Further, in Africa which has a high incidence of HIV/AIDS, the
provision of irradiated, low microbial load, ready-to-eat foods
could be of great benefit to these immuno- depressed patients
(Duodua et al., 1999).
9.1. Radiation sources used for food irradiation
Expert Committee of WHO/IAEA/FAO in accordance
with international regulations of Codex General Standards for
food irradiation approved three types of ionizing radiation for
treating foods as reported by WHO ( 1988).
a) Electron beams: which are negatively charged particles,
produced from electron accelerator machine sources
operated at or below an energy level of 10 MeV.
b) X-rays: which are electromagnetic radiations of short
wave length, generated from machine sources operated at
or below an energy level of 5 MeV.
c) Gamma rays: which are electromagnetic radiations of
very short wavelength emitted by the nuclei of
radioactive substance during decay from radioisotopes
mainly Cobalt-60 and Cesium-137.
9.2. Wholesomeness and safety aspects
No other method of food processing has been subjected
to such a thorough assessment of safety as the radiation
processing. The various aspects of wholesomeness and safety of
radiation
33
Introduction
Review
of Literature
processed foods have been studied in great detail (WHO, 1994;

Diehl, 1995). These include:
Possibilityof induced radioactivity Microbiological safety
Nutritional adequacy
Animal feeding
Safety of chemical changes
Human trials
At the energies of the gamma rays from Cobalt-60 (1.3
MeV) and those recommended for X-rays (5 MeV) and
accelerated electrons (10 MeV), no induced radioactivity has
been observed. The microbiological aspects of radiationprocessed foods have been studied in detail. None of these
studies have indicated that foods preserved by radiation pose
any special problems in relation to microflora. It has been found
that there are no unique radiolytic products formed and free
radicals in the system disappear depending on the nature of the
commodity and its post-irradiation storage and treatment. In
fact, the chemical differences between radiation processed foods
and non-irradiated foods are too small to be detected easily.
Though rough composition of food remains largely unchanged,
some losses in vitamins may be encountered. However, these
losses are often minor and could be made up from other sources.
9.3. International approval
Stevenson (1994) reported that irradiation technology is
the answer to many questions, since it creates the possibility of
putting foods on the market that are glowing with freshness,
and exempt of pathogens. This technology was approved in
1981 by the FAO/IAEA/WHO joint Committee on the
Wholesomeness of Irradiated Food. It was stated that,
irradiation of food at doses up to 10 kGy introduced no special
nutritional problem and irradiated food does not need any
toxicological tests. In, 1997 an FAO/IAEA/ WHO study group
on high dose irradiation examined
34
Introduction
Review
of Literature
the safety studies carried out on food irradiated with doses

higher than 10 kGy and concluded that food irradiation at any
dose appreciate to achieve the intended technological objectives
is both safe to consumer ( WHO, 1997).
In 1999, a coalition of U.S. food processors petitioned
the U.S. Food and Drug Administration to amend U.S.
regulations to allow doses of up to 4.5 kGy for a wide variety of
refrigerated and ready to eat meat and vegetable products,
including juices, with doses up to 10.0 kGy sought for frozen
meat, vegetable, and juice products, elimination of human
pathogens is the primary goal of these requested dose limits; the
potential for extension of shelf life is regarded as a secondary
goal ( Anonymous, 2000a) Therefore, as scientific data
accumulate and commercial interest in irradiation increases,
U.S. and international regulations concerning the irradiation of
fresh produce and juices are expected to change to reflect
increased understanding. Irradiation as a food treatment is
endorsed by a variety of professional and governmental
organizations such as the United States Food and Drug
Administration ( Anonymous, 2000 b), U.S. General
Accounting Office ( Anonymous, 2000a ), American Dietetic
Association ( Wood and Bruhn, 1999; WHO , 1999) .
Aymerich et al. (2008) reported that all irradiated foods
are required to have a label indicating that they have received
such treatment. Irradiation technology was promoted by the
FAO in the Codex Alimentarius in 2003 and has been well
accepted in 50 countries.
Farkas and Mohacsi-Farkas (2010): stated that the safety of
consumption and wholesomeness of irradiated food have been
extensively studied in international cooperations. Numerous
international expert groups set up jointly by the FAO, the IAEA
and the WHO, or the Scientific Committee on Food of the
35
Introduction
Review
of Literature
European Commission concluded that foods irradiated with

appropriate technologies are both safe and nutritionally
adequate. A Codex General Standard for Irradiated Foods and a
Recommended International Code of Practice for Radiation
Processing of Food have been developed. Specific applications
of food irradiation are approved by national legislations in over
55 countries worldwide among of which are the USA, Egypt,
and China and across Latin America
9.4. Technological benefits and advantages
Major technological benefits that can be achieved by
radiation processing of food include:
Disinfestations of insect pests in stored products.
Inhibition of sprouting in tubers, bulbs and rhizomes.
Delay in ripening and senescence in fresh fruits and
vegetables.
Destruction of microbes responsible for food spoilage
including fresh meat, poultry, fish, fruits, vegetables and
their products.
Elimination of parasites and pathogens of public health
importance in food on the basis of dose requirements. These
benefits could be classified in to low dose, medium dose, and
high dose applications (Dris and Jain, 2004).
The technology can be applied in three broad areas
For strengthening food security.
For improving food safety.
For increasing international trade.
36
Introduction
Review
of Literature
It can improve food security by cutting down food losses

caused by storage insects, microorganisms, and physiological
changes. It is believed that 2050% of the agricultural produce
is lost due to inadequate post-harvest practices. It can help
reduce the risk of food borne-illnesses by eliminating pathogens
and parasites in food, thus improving food safety. It can also
help to boost international trade by overcoming quarantine
barriers and by improving quality and marketability of
agricultural products.
Radiation technology offers several advantages for
processing food. These advantages are listed below:
It is a physical, non-additive process, causing minimal change
in food.
It is highly effective compared to chemicals and fumigants.
It does not leave harmful residue in food.
It can be applied to bulk as well as pre-packaged food.
It is a cold process and preserves food in natural form.
It does not destroy heat-labile aroma constituents of food.
The process is safe to workers and friendly to environment
(Dris and Jain, 2004).
9.5. Limitations
The technology has also a few limitations. These are:
The technology cannot be applied indiscriminately to all foods.
The technology is also not appropriate for destruction of
viruses and enzymes. Therefore, to meet such an objective the
technology should be complemented with other methods such as
mild heat.
37
Introduction
Review
of Literature
Similarly, a food product which is molded and already

contaminated with preformed microbial toxins can not be
treated with radiation as a part of the good irradiation practice.
Some of these measures are part of the good manufacturing
practices for any food processing method.
The technology is also capital intensive.
In case of gamma irradiation plants needs handling of
radioisotopes requiring proper safeguards and approvals (Dris
and Jain, 2004).
10. Irradiation effects
Generally, ionizing radiation has been found to be very
effective in inactivating microbial cells. The mechanism of
microbial death is a sequence of the ionizing action of highenergy radiation. Most studies indicated that the damage of the
microbial cell DNA, which may result in loss of ability to
reproduce, is a primary cause of lethality but damage of other
sensitive molecules (cell membranes, intracellular constituents
of biological function importance) may also have effects.
Ionizing radiation can affect microbial cell DNA by two
mechanisms, either directly by high-radiation energy which
cause direct damage to DNA or indirectly through the effect of
primary water free radicals (H, OH, e) which are formed
upon water radiolysis of the microbial cells and water
surrounding cells (Ingram and Roberts, 1980). The OH
radical is the most important oxidizing agent affecting microbial
cell. Therefore, it is expected that microorganisms are more
resistant to radiation in the dry state than in the presence of
water.
38
Introduction
Review
of Literature
10.1. Relative radiation resistance of microorganisms

Microorganisms differ greatly in their resistance to
ionizing radiation. The radiation resistance of microorganisms is
measured by the so called radiation reduction decimal dose
(D10-value). D10-value, for a microbe is defined as the radiation
dose (kGy) required to reduce the number of that microbe by a
10-Log cycle or to kill 90% of the numbers (Ingram and
Roberts, 1980). The sensitivity of microorganisms to ionizing
radiation is often compared on the basis of D10-values. To
accommodate non-liner survivor curves, the so-called (most
probable effective dose) to achieve n Log cycles reduction
( MPED )n has been introduced ( Mossel and Degroot, 1965
).For all practical purposes the MPED is the dose of irradiation
that, at a given level of initial contamination reduces the final
cfu count to a non-detectable level, i.e. no more than 101 - 102
cfus per gram of product.
Generally, the radioresistance of the bacterial species
were arranged in the following order : Pseudomonas, Coliform,
Staphylococci, Salmonella , Streptococci, sporeformer and
finally Clostridium botulinum type A and B ( Thornley, 1963 ).
Christensen (1970) arranged the large groups of
microorganisms according to increasing radioresistance in the
following order: gram-negative bacteria, vegetative forms of
gram-positive bacteria, mold, yeast, bacterial spores and viruses.
Ingram (1975) reported that the gram negative bacteria
including the common spoilage organisms of many foods, e.g.
Pseudomonas and particularly certain species of pathogenic, e.g.
Salmonella and Shigella were more sensitive than vegetative
gram-positive bacteria. Rowley et al. (1978) concluded that
bacterial spores are the most radiation resistant and Gramnegative rods are the most sensitive. Moraxella-Acinetobacter,
Micrococcus radiodurans,
39
Introduction
Review
of Literature
Pseudomonas radiora and Deinococcus spp. are able to survive

high irradiation doses (Diehl, 1995).
Sommer (1973) indicated that dose response curves of
sexual spores of important fungal genera showed a wide
spectrum of radiation resistance when suspended in water or
buffer and irradiation in presence of oxygen. He indicated that
the D10 values for penicillium expansum, Mucor sp., Botrytis
cinerae, Rhizopous stolonifes, Alternaria citri, Cladosporium
herbarum and Alternaria tenuis were approximately 0.5, 0.6,
0.8, 0.9, 1.0, 1.1 and 1.4 KGy, respectively. O'Neill et al.
(1991) reported that species of Fusarium and Alternaria were
more resistant to irradiation than Aspergillus spp. and
Penicillium spp. However, radiation doses required for direct
suppression of postharvest pathogens are generally above the
tolerance level of the fruit and result in radiation damage
(Barkai-Golan, 1992).
D10values have been reported for yeasts and molds in
the range of 1 to 3 kGy (Narvaiz et al., 1992). Kim et al.
(2007) tested the radiation-resistant of bacteria N-1(Bacillus
megaterium IAM 13418T) and N-2 (Exiguobacterium
acetylicum X70313) which were isolated from the kale juice.
From the results of the inoculation test, it is suggested that even
though the radiation-resistant microbes in the kale juice
survived gamma irradiation, the 35 kGy radiation doses may
prevent a follow up microbial growth during a 3 days postirradiation storage at 10C. Monk et al. (1994) reported that the
main effective microorganisms in juice irradiation were yeasts
and molds, which have a higher D10- value than bacterial
pathogens. In fruit juices, D10- values have been reported for
yeasts and molds are in the range of 1-3 kGy (Narvaiz et al.,
1992) and 0.3-0.7 kGy for pathogenic bacteria (Buchanan et
al., 1998, Niemira et al., 2001). Buchanan et al. (1998) showed
that a dose of 1.8 kGy should be sufficient to
40
Introduction
Review
of Literature
achieve the 5D-inactivation of E.coli recommended by the

National Advisory Committee for Microbiological Criteria For
Food Irradiation.
10.2. Effect of irradiation on microbial load of fresh fruits
and vegetables
Generally, irradiation has been found to be effective in
reducing the microbial load of fresh fruits, vegetables and their
minimally processed produce. Several researches have reported
that a low-dose irradiation can improve the microbial quality of
various fresh-cut vegetables such as shredded and cut- carrots
(Hagenmaier and Baker, 1998), cut lettuce (Hammad et al.,
2010), diced celery (Prakash et al ., 2000), green onion leaves
(Fan et al., 2003a) coriander leaves (Kamat et al ., 2003).
Hammad et al. (1995) found that application of gamma
irradiation at 1 and 2 kGy almost inactivated Gram-negative
bacteria. Pseudomonas and Enterobacteriaceae and caused
great reduction in total aerobic bacterial counts, molds and
yeasts of fresh strawberries. These doses extended the
refrigeration (4C1) shelf-life of strawberries to 18 and 21
days, respectively against only 10 days for non-irradiated ones.
Howard et al. (1995) reported that 1.0 kGy reduced total
aerobic bacteria and lactic acid bacteria in Pico de gallo, a cold
salad made of tomatoes, yellow onions, and jalapeno peppers,
over six weeks of refrigerated storage, thereby extending the
shelf life. Farkas et al. (1997) showed that ionizing radiation at
1 kGy reduced loads of bacteria, improved microbiological shelf
life, and extended sensory quality of precut peppers and carrots.
Prakash et al. (2002) indicated that irradiation at 3.70
kGy resulted in no aerobic populations through day 12 and
significantly fewer colonies through day 15 in diced Roma
tomatoes. Where, Zhang et al. (2006) reported that the the
41
Introduction
Review
of Literature
number of aerobic mesophilic bacteria on fresh-cut lettuce

irradiated with 1.0 kGy was reduced by 2.35 logs and sensory
quality was maintained best during storage for 8 days at 4 C.
Hussain et al. (2008) reported that the gamma-irradiation dose
range of 1.21.4 kGy significantly (p< 0.05) delayed the
decaying of the peach fruit by 6 days under ambient conditions
and by 20 days under refrigerated storage conditions. Wani et
al. (2008) reported that yeast and mold count of pears was
markedly reduced by both irradiation and low-temperature
storage. No yeast and mold count was recorded in samples
irradiated at dose beyond 1.4 kGy after 7 days of ambient
storage. As the storage period advanced, yeast and mold count
increased and was significantly (p<0.05) higher in unirradiated
samples under both the storage conditions. Hammad et al.
(2010) found that irradiation caused a great reduction in all
microbial loads of fresh cut lettuce and during refrigerated
storage, the counts of all microorganisms increased, but the rate
of increase was slower in irradiated samples in comparison with
unirradiated ones. Irradiation dose of 3 kGy was the optimum
dose for preservation of fresh cut lettuce which extended the
refrigeration shelf-life up to 20 days and it was sufficient in
eliminating all pathogenic bacteria without adverse effect on
sensorial quality and slight effect on chemical quality.
10.3. Effect of irradiation on foodborne pathogenic bacteria
contaminating fresh fruits and vegetables and their
minimally processed produce
The preparation and distribution of minimally processed
vegetables is a rapidly developing industry that provides the
consumer with convenient and nutritious food. However,
minimally processed vegetables may undergo substantial
mechanical injury due to cutting with the result that
42
Introduction
Review
of Literature
microorganism can grow rapidly upon exposure to nutrient

juices, thus leading to elevated respiration, rapid deterioration
(Saltveit, 1997) and insecurity of this product. Many
investigators have shown that, irradiation approved to be a
highly effective method for eliminating human food borne
pathogens on and in fresh fruits, fruit juices, fresh vegetables,
vegetables juices, fresh cut vegetables, vegetable salads and
seed sprouts with maintenance of their freshness and quality
attributes (Chervin and Boisseau, 1994; Prakash et al., 2000
and Abu- El Nour, 2007). Farkas et al. (1997) found that dose
of 1.0 kGy reduced total aerobic plate count and Listeria
monocytogenes by ~4 logs (99.99%) on pre-cut bell pepper;
however, on peppers stored at 15 or 10C, the pathogen re-grew
to initial levels within 4 days, while pathogen levels remained
low on peppers stored at 5C. Also in that study, spoilage
bacteria were reduced by 5 logs on carrot cubes by 1.0 kGy. The
authors concluded that irradiation, when combined with good
manufacturing practices, could effectively reduce pathogen
levels throughout the useful shelf life of the produce.
Martins et al. (2004) reported that low-dose irradiation
treatments (< 2 kGy) effectively inactivate most of the food
borne pathogens including Escherichia coli O157:H7, Yersinia
and Salmonella with little or no defects on the sensorial quality
of minimally processed vegetables. Lee et al. (2006) found that
all the bacterial contents of test pathogens into the samples of
cucumber blanched and seasoned spinach, and seasoned
burdock were reduced to below the limit of detection by 3 kGy
irradiation. Todoriki et al. (2009) reported that a 1.25 kGy dose
of gamma irradiation was found to be sufficient to eliminate
Listeria monocytogenes on whole cherry tomatoes.
43
Introduction
Review
of Literature
10.4. Effect of irradiation on microbial quality of fruits and

vegetables juices
The value of fresh fruits and vegetable juices lies in the
primary characteristics of freshness and convenience. The
growing demand for fresh fruits and vegetable juices is
stimulating the production of vegetable juice type products in
the food industry. However, the development and marketing of
these juices are limited because they are highly perishable and
have short shelf-life due to the growth of putrefying
microorganisms (Song et al., 2007). Thus, fresh fruits and
vegetable juices should be consumed within 24 h in general. The
microbial flora in the fruit and vegetable juices is thus likely to
be similar to the flora in raw fruits and vegetables. Bacteria in
fresh vegetable juice, especially pathogens that come from soil,
water or the hands of workers, affect not only the public health,
but also the product quality. Therefore, vegetable juice should
undergo some type of processing to inactivate most of the
microorganisms. Recent regulations by the FDA have required
processors to achieve a 5 log reduction in the numbers of the
most resistant pathogens in their finished products. This rule
comes after a rise in the number of food borne illness outbreaks
and consumer illnesses associated with consumption of
untreated juice products. The ruling has accelerated the search
for novel non thermal processes that can ensure product safety
yet maintain the desired nutritional and sensory characteristics
(Tiwari et al., 2009).
Buchanan et al. (1998) showed that irradiation (1.0kGy)
effectively inactivated Escherichia coli O157:H7 in inoculated
commercial apple juices, with D10 values of 0.12, 0.16, and 0.21
kGy for the three isolates tested. Niemira et al. (2001) found
that the resistance of four Salmonella isolates irradiated in
orange juice also varied, with D10 values of 0.71 kGy ( S.
anatum ), 0.48
44
Introduction
Review
of Literature
kGy ( S. Newport ), 0.35 kGy ( S. infantis ), and 0.38 kGy ( S.

Stanley ). For the most resistant isolate (S. anatum), 3.5 kGy
was indicated as the dose required to achieve a 5- log reduction.
Pickett et al. (2000) also found that Salmonella hartford was
reduced to undetectable levels in irradiated (3.0 kGy) orange
juice.
Song et al. (2007) emphasized that all the aerobic and
coliform bacteria in the carrot juice were eliminated by
irradiation at a dose of 3 kGy, whereas about 102 CFU/ml of the
bacteria survived in the kale juice irradiated at up to 5 kGy.
However, the cells that survived from irradiation in the kale
juice did not grow, whereas those of the non-irradiated samples
reached 10 9 CFU/ml after 3 days of storage at 10 C.
Application of gamma radiation to pomegranate juice
(Alighourchi et al., 2008), carrot and kale juice (Kim et al.,
2007) and UV radiation to orange, guava and pineapple juice
(Keyser et al., 2008) has been reported for the inactivation of
both spoilage and pathogenic microorganisms thereby extending
their shelf-life and ensure safety.
Song et al. (2007) indicated that irradiation at 35 kGy
improved the microbial shelf-stability of the juices (carrot and
kale juices), and prolonged their shelf life up to 3 days at 10C
whereas the shelf life of the non-irradiated control was limited
to 1 day. So, it was considered that gamma irradiation was not
only effective in pasteurization of fresh vegetable juices but also
in prolonging the shelf life.
10.5. Effect of irradiation on the main chemical components
of fresh fruits and vegetables and their produce
10.5.1. Anthocyanin
Anthocyanins are plant pigments responsible for the
orange, red and blue colours of various fruits and vegetables
45
Introduction
Review
of Literature
(Rentzsch et al., 2007) Irradiation effects on anthocyanin

pigments depend upon the nature of anthocyanin for example;
diglycosides are relatively stable towards irradiation dose
compared to monoglycosides. Chachin and Ogata (1969) have
previously investigated the effect of different irradiation doses
of gamma radiation on grape, apple and orange juices
anthocyanins. They found that irradiation at 10 kGy caused
evident loss of grape juice anthocyanins. In contrast, Ayed et al.
(1999) reported that the anthocyanin content in grape pomace
increasesd with irradiation doses (0-9 kGy), especially at 6 kGy.
This increase in the measured content may be due to the
extraction of bound pigments by degradation of the cell wall.
Bakowska et al. (2003) reported a strong negative
influence of UV irradiation on the complex of cyanidin-3glucoside with co-pigment compared to thermal treatment at 80
C. However, the presence of certain copigments can inhibit the

degradation effect of UV on anthocyanins improving the
cyanidine-copigment complex. Anthocyanins exposure to UV-C
(0.56-13.62 kJ/m2) showed insignificant changes in the
anthocyanins as well as the antioxidant capacity in pomegranate
arils (Lopez-Rubira et al., 2005). Alighourchi et al. (2008)
studied the effect of gamma irradiation (0-10 kGy) on the
stability of anthocyanins in pomegranate juice. Their results
indicated that the irradiation at all applied doses, significantly
reduced total and individual anthocyanins. They added that
irradiation with higher dosages (3.5-10 kGy) had undesirable
effect on the total content of anthocyanins. Erkan et al. (2008)
reported that UV-C treatment for different durations (1, 5, and
10 min) increased the antioxidant capacity and the
concentrations of anthocyanins and phenolic compounds in
strawberries.
46
Introduction
Review
of Literature
10.5.2. Lycopene
Lycopene is a vibrant red carotenoid that serves as an
intermediate for the biosynthesis of other carotenoids (Stahl
and Sies, 1996). To our knowledge, there is no information
about the effect of gamma irradiation on lycopene content of
major varities of fruits and vegetables.
Arts-Hernandez et al. ( 2010 ) studied the effects of
four pre-packaging UV-C illumination doses (1.6, 2.8, 4.8 and
7.2 kJm2) on quality changes of watermelon cubes stored up to
11 days at 5 C. Control cubes showed a 16% decrease in
lycopene content after 11 days at 5 C similar to that found for
the high UV-C treatment. However low UV-C treated
watermelon cubes preserved their initial lycopene content (2.8
kJm2) or it was slightly decreased (1.6 kJm2).
10.5.3. Carotenoids
Carotenoids are among the most important nutrients in
food, owing to their diverse functions and actions. The attractive
red or yellow colour conferred to many foods is their most
apparent contribution to food quality.
Beta-carotene is a natural compound that is omnipresent
in yellow and green vegetables and fruits, such as carrot,
spinach, sweet potato, pumpkin, broccoli, peppers, pink
grapefruit, papaya and peach. The presence of light, acids, heat,
oxygen, the methods of preservation and the storage time are
destruction factors which lower the total -carotene level and
may cause isomerisation reactions. Chachin and Ogata (1969)
treated grape, apple and orange juices with different irradiation
doses of gamma radiation. Loss of orange juice beta carotene
was evident after 10 kGy and
47
Introduction
Review
of Literature
was dose dependent up to 80 kGy. Josimovic (1983) did not

notice any significantly quantitative changes in some chemical
compounds of dehydrated parsley irradiated with doses up to 20
kGy.
According to the International Consultative Group on
Food Irradiation (ICGFI), gamma-irradiation does not affect the
content of -carotene or other carotenoids with pro-vitaminic A
activity (ICGFI, 1999). Gamma irradiation doses (0, 10, 20, 30
kGy) induced a slight decrease in -carotene content in
artichoke (Koseki et al., 2002) Fresh-cut mangoes UV-C
irradiated for 0, 10, 20, and 30 min, showed decrease in carotene content (Gonzalez-Aguilar et al., 2007). Hajare et al.
( 2007 ) reported that total carotenoids content of pea sprouts
stored at 4C, as well as at 8C, for 12 days remained almost
unchanged after irradiation (1 and 2 kGy ) as well as during
storage.
10.5.4. Ascorbic acid
Vitamin C is the most important vitamin for human
nutrition that is supplied by fruits and vegetables and their
juices. Since vegetables are an important source of vitamin C in
the total diet, it is important that any preservation method used
to extend their shelf life will not markedly affect the vitamin C
content. It is well known that vitamin C is the most sensitive of
all water-soluble vitamins to an irradiation (Kilcast, 1994).
However, it has been noted that when reporting vitamin C levels
in irradiated food, many workers have not taken into
consideration the fact that ionizing radiation can cause a partial
conversion of ascorbic acid to dehydroascorbic acid. It is also
possible that some degradation of the vitamin could have
occurred through its attack by free radicals. The amount of
vitamin loss due to food irradiation is affected by several
factors, including dose, temperature, presence of oxygen, and
food type. Generally, radiation at low
48
Introduction
Review
of Literature
temperatures in the absence of oxygen reduces any vitamin loss

in foods, and storage of irradiated foods at low temperatures
also helps prevent future vitamin loss (Olson, 1998).
Mahmoud (1973) reported that exposing of onion to
gamma rays increased ascorbic acid percentage. On the other
hand, Hegazi (1981) studied the effect of gamma irradiation on
ascorbic acid percentage of strawberry and he observed that
irradiation treatment reduced ascorbic acid content. While,
Orabi (1981) found no significant effect on the concentration of
vitamin C in potato tubers due to gamma irradiation by doses of
4 and 8 Gy. However, Singh (1990) found that the exposure of
mangoes to gamma irradiation significantly decreased the
vitamin C in a few varieties of mangoes, while in the others, the
vitamin C level was unaffected by radiation.
Mitchell et al (1992) reported that irradiation doses at 6
kGy did not result in a significant decrease in total ascorbic acid
(vitamin C) and caused an increase in dehydro-ascorbic acid
content in mango. Lacroix and Gagnon (1993) found that
mangoes exposed to gamma rays at 0.49 to 0.77 kGy was
slightly had increasing in ascorbic acid content. In contrast,
Shengfu et al. (1993) stated that kiwi fruits irradiated with
gamma rays at dose of 0.6 KGy and then stored in air ventilated
room had slight decrease in vitamin C content. Ibrahim (1996)
stated that gamma irradiation treatments (1, 2 and 4 kGy)
reduced the ascorbic acid content in carrot roots. The higher the
irradiation doses the lower the ascorbic acid content. While,
Graham and Stevenson (1997) noticed that there was no effect
of gamma irradiation on ascorbic acid content in strawberry,
Ladaniya et al (2003) found that exposing of three species of
citrus to gamma rays up to 1.5 kGy decreased vitamin C
content. Patil et al (2004) reported that irradiation doses of
gamma rays up to 0.7 kGy had no remarkable
49
Introduction
Review
of Literature
effect on vitamin C content at early season of grapefruit. Kim

and Yook (2009) demonstrated that irradiation of kiwifruit up
to 3 kGy had negative effects on vitamin C content and
antioxidant activity. Arts-Hernandez et al. (2010) stated that
UV-C radiation did not significantly affect the vitamin C
content in watermelon cubes.
10.6. Effect of irradiation on sensory properties
Sensory evaluation is defined as a scientific method used
to evoke, measure, analyze, and interpret those responses to
products as perceived through the senses of sight, smell, touch,
taste, and hearing ( Lawless and Heymann, 1999).
Depending upon the research question, sensory food
science also utilises physicochemical, physiological, and
consumer-based research methods. The importance of sensory
food science is based on the relevance of consumer perceptions
to the acceptance and commercial success of foods and
beverages and on the significance of food for human well-being
and health. In food companies, sensory food science can be of
great value to both tactical and strategic research goals. Human
senses, in particular the chemical senses, are tuned to act as
composite gatekeepers for food intake. This biological function
protects us from eating spoiled or otherwise unfit items and
encourages eating nutritious or otherwise beneficial items
(Breslin and Spector, 2008).
Irradiation induces negligible or subtle losses of
nutrients and sensory qualities in food compared to thermal
processing as it does not substantially raise the temperature of
food during processing (Wood and Bruhn, 2000). Chachin
and Ogata (1969) treated grape, apple and orange juices with
different doses of gamma radiation. Browning was reported in
apple juice after 5
50
Introduction
Review
of Literature
kGy. High dose irradiation (10 kGy) of orange juice resulted in

a similarly unacceptable degree of flavor degradation and
browning. Abdollaoui et al. (1995) observed that according to
the sensorial evaluators, the taste of the irradiated fruits is
sweeter than in non-irradiated fruits. Spoto et al. ( 1997 )
reported that the highest dose ( up to 5.0 kGy ) combined with
25C storage resulted in loss of orange flavor and an
increase in bitter , medicinal , and cooked ratings by
sensory panelists. They added that lower dose (2.5 kGy) and
cooler storage (0 or 5C) were proposed as an acceptable
processing regimen.
Pickett et al. (2000) reported increasing off- flavor in
unpasteurized orange juice irradiated to 3.0 kGy, rendering the
juice unpalatable. Niemira et al. (2001) found no evident
changes in the appearance or aroma of reconstituted orange
juice irradiated to 2.5 kGy. Pasteurized orange juice irradiated to
doses as high as 5.0 kGy demonstrated little reduction in
product color or concentration of key aroma and flavor
compounds. Unpasteurized apple cider irradiated to 3.0 kGy
was identifiable, although not unacceptable, to sensory panelists
evaluating aroma; similar results were obtained with
reconstituted and fresh orange juices.
Song et al. (2007) postulated that the sensory quality of
the non-irradiated carrot juice decreased with the storage time,
and it appeared to be the worst at storage of day 3. Panelists
evaluated that the control could not be used for food after 2- day
storage because of the deterioration in quality by spoilage. In the
observation of the appearance, the color of the juice showed a
critical change (P < 0.05). The color of the irradiated samples
maintained its original color over time, whereas the color of the
control samples became darker. This result may be due to a
decrease of the polyphenol oxidase activity by irradiation
51
Introduction
Review
of Literature
(Hanotel et al., 1995), which is responsible for the browning of

fruit and vegetables. Thus, an important improvement of the
sensory parameters is achieved by employing the irradiation
treatment. Song et al. (2007) found that there was no difference
in sensory evaluation (odor, color, taste and overall
acceptability) results of carrot and kale juices immediately after
irradiation at 3 kGy. At day 3 storage (10C), the sensory
quality of the irradiated juice was adequate, while the quality of
the non-irradiated control was deteriorated.
10.7. Viscosity effects
The viscosity of fluid food is an important property
which has many applications in food technology, such as
developing food processes and processing equipment, the
control of products, filters and mixers, quality evaluation and an
understanding of the structure of food and raw agricultural
materials (Alvarado and Romero, 1989).Viscosity is an
important quality attribute limiting the consumer acceptability
of fruit juices that can be determined by factors such as the
cultivar of fruits or the maturity of the fruit at the moment of
processing (Tiziani and Vodovotz, 2005).
Consistency of tomato products refers to their viscosity
and the ability of their solid portion to remain in suspension
throughout the shelf-life of the product. The consistency of
tomato products is strongly affected by the composition of the
pectins. Controlling the breakdown or retention of the pectins,
and the enzymes that lead to changes in the pectins, is thus of
great importance during processing (Hayes et al., 1998). Two
enzymes, pectin methylesterase (PME) and polygalacturonase
(PG) are involved in the breakdown of pectins. The action of
PME makes the pectin susceptible to further degradation by PG
because this enzyme acts only on segments of the pectin chain
52
Introduction
Review
of Literature
that have been demethylated by PME. The degradation of the

pectin chains reduces the viscosity of the juice Crelier et al.
(2001). Therefore, experimental measurements of viscosity are
necessary for the characterization of fluid foods (Juszczak and
Fortuna, 2004).
Many food products such as sauces, syrups, ice cream,
instant foods, beverages and confectionaries, marshmallows and
candies contain hydrocolloids in their formulations (Kayacier
and Dogan, 2006). Exposure to irradiation may produce certain
changes in the chemical properties of foods. Thus, the use of
irradiation in food hydrocolloids may cause a change in the
consistency and viscosity characteristics. As a result, the final
product containing irradiated hydrocolloid may be inversely
affected by the irradiation dose. Gagnon et al. (1968) showed
that irradiation lower the viscosity of apple juice and therefore
increases rate of filtration in it. Hayashi and Todoriki (1996)
found that viscosity values of irradiated pepper samples at 5
kGy were lower than 170mPa.s, while those of unirradiated ones
were higher than 300 mPa.s. Swailam (1998) detected a
reduction in the consistency index of the mango pulp, and he
reported that it may be taken in consideration for the detection
of irradiated mango pulp at zero time and during storage.
11. Preservation of juices by irradiation and other
technologies
Gamma irradiation can be used for many applications
related to food science. Low dose of irradiation can be used to
prolong the shelf life of many fruits and vegetables by reducing
microbial spoilage, reducing the rate of respiration and delayed
ripening. The use of preservatives in combination with
irradiation, especially natural preservatives or natural active
edible coating, represents a challenge for scientists to develop
53
Introduction
Review
of Literature
new food preservation methods to improve the shelf-life

without affecting the sensorial quality. Some studies available
have demonstrated synergistic effects to reduce the content of
microorganisms in fruit (Vachon et al., 2002) and vegetables
(Lafortune and Lacroix, 2004). It has also been demonstrated
that the active antimicrobial compounds present in natural
preservatives can improve the radiosensitivity of bacteria
(Mahrour et al., 2003; Chiasson et al., 2004).
When irradiation is used in combination with other
technologies such as mild heat, addition of natural preservatives,
plant extracts and refrigeration storage, the global efficiency is
reinforced through synergistic action, and the irradiation dose
can be reduced without affecting the product quality (Farkas,
1990). Tiwari et al. (2009) reported that in response to
consumer demand for greener additives, combinations of
various extracts from natural plants or antimicrobials agents can
be explored further to provide improved stability of
anthocyanins and polyphenols through co pigmentation while at
the same time providing synergy for microbial inactivation. The
applications of natural antimicrobial agents are likely to grow
steadily in the future because of greater consumer demands for
minimally processed foods and those containing naturally
derived preservation ingredients. More complex considerations
arise for combinations of technologies, particularly with respect
to optimisation of practical applications
11.1. Irradiation in combination with refrigeration
Studies have shown that gamma irradiation alone and
combined with other treatments (for example: refrigeration)
decreased the microbial contamination level, thereby leading to
an enhancement of the shelf-life (Lacroix et al., 1991).
54
Introduction
Review
of Literature
Lacroix and Ouattara (2000) reported that strawberries,

for instance, may tolerate doses as high as 3.0 kGy. At this dose,
the infection of fruits is eliminated and the quality of the
strawberries is untouched for a period of 14 days if kept at 5C.
Song et al. ( 2007 ) indicated that irradiation at 3-5 kGy
improved the microbial shelf-stability of the juices ( carrot and
kale juice ), and prolonged their shelf life up to 3 days at 10C
whereas the shelf life of the non-irradiated control was limited
to 1 day. Alighourchi et al. (2008) reported that irradiation at
0.5 and 2.0 kGy reduced the growth rate of bacteria and fungi of
the selected pomegranate juices during the first 3 days of
storage at 4C. Hussain et al. (2008) reported that the firmness
of unirradiated control samples of peach fruit decreased from an
initial value of 9.4 kg at harvest to 1.1 kg after 9 days of
ambient storage and 1.2 kg after 20 days of refrigerated storage
(31 C), firmness of irradiated samples (range 1.0-1.4 kGy)
decreased to 1.3 kg after 12 days of ambient storage and 1.5-1.9
kg after 20 days of refrigerated storage. It is also observed that
irradiation significantly (p<0.05) delayed the delaying of
peaches under both storage conditions. Under ambient storage
conditions, unirradiated samples were found to have started
decaying after 3 days of storage, where as no decay was
observed in irradiated samples over the same storage period.
Irradiated samples except those irradiated in the dose range of
1.0-1.4 kGy started decaying after 6 days of storage, while
under refrigerated storage no decay was observed up to 20 days
in sample irradiated to 1.0-1.4 kGy dose. Wani et al. (2008)
reveals that gamma irradiation alone and in combination with
refrigeration (31C) proved significantly effective in extending
the storage life of pear. Dose range of 1.5-1.7 kGy resulted in
shelf-life extension of the fruit by 14 days under ambient
storage conditions. Irradiation in combination with refrigeration
retarded the onset of decay by 45 days as against the 35% decay
in unirradiated control samples. Dose range of 1.5-1.7
55
Introduction
Review
of Literature
kGy gave a further extension of 8 and 4 days during additional

ambient storage of pears following 30 and 45 days of
refrigeration (31C), respectively.
11.2. Irradiation in combination with natural additives
Many spices, condiments, and plant extracts have strong
medicinal, preservative, and antioxidant properties (Zaika,
1988). The antimicrobial activity of these ingredients is
attributed to their essential oils, which are lipophilic and
penetrate through the membrane to the interior of the cell and
perform the inhibitory activity at the target site (Smith-palmer
et al., 1998). Cinnamon effectively inhibits the growth of
bacteria (with gram- positive being more sensitive than gram
negative), yeasts, and molds (Shelef, 1983). The major
antimicrobial components of spices and their essential oils are,
for example, eugenol in cloves, allicin in garlic, cinnamic
aldehyde and eugenol in cinnamon, carvacrol and thymol in
oregano and thyme, and vanillin in vanilla beans.
The antimicrobial activity of some essential oil
components against foodborne pathogens, including mycotoxinproducing fungi, has also been tested (Bullerman, 1977).
Cinnamon oil and clove oil are both natural preservative and
flavouring substances that are not harmful when consumed in
food products. There have been a number of reports of
substances in each of cinnamon and clove oils that inhibit the
growth of molds, yeasts and bacteria. Both cinnamon oil and
clove oil added at 2% in potato dextrose agar (PDA) completely
inhibited the growth of seven mycotoxigenic molds (Aspergillus
flavus, A. parasiticus, A. ochraceus, Penicillium sp., P.
roqueforti, P. patulum, and P. citrinum) for various times up to
21 days (Azzouz and Bullerman, 1982) and could also inhibit
the growth of yeasts (Conner and Beuchat, 1984).
56
Introduction
Review
of Literature
Smith-Palmer et al. (1998) found that the minimum

inhibitory concentrations (MIC )of cinnamon and clove essential
oils against Campylobacter jejuni, Escherichia coli, Salmonella
Enteritidis, Listeria monocytogenes, and Staphylococcus aureus
were 0.05, 0.04-0.05, 0.04-0.05, 0.03, and 0.04%, respectively,
in an agar dilution assay. Smid and Gorris (1999) reported that
cinnamic aldehyde inhibited growth of both bacteria and fungi
and increased shelf life of treated packaged tomatoes. Soliman
and Badeaa (2002) found that <500 ppm of cinnamon oil can
inhibit A. flavus, A. parasiticus, A. ochraceus and Fusarium
moniliforme on potato dextrose agar.
The greater effectiveness of the cinnamon bark oil on
Salmonella Enteritidis and Escherichia coli O157:H7 in fruit
juices was associated to the media pH, being more effective in
media with lower pH. Burt (2004) reported that the bacterial
susceptibility to the antimicrobial effect of essential oils appears
to increase with a decrease in the pH of the food, because at low
pH the hydrophobicity of an essential oil increases, enabling it
to more easily dissolve in the lipids of the cell membrane of
target bacteria.
Although, the mechanism of action of cinnamon oil on
the microbial cells is still unclear, Wendakoon and Sakaguchi
(1995) and Burt (2004) have indicated that the interaction of
carbonyl group of the cinnamaldehyde, main compound of
cinnamon oil from bark, on the cell proteins embedded in the
cytoplasmatic membrane appear to inhibit the action of the
enzymes amino aciddecarboxylases, which are necessary for the
amino acids biosynthesis and biodegradation. Oussalah et al.
(2006) reported a release of the cell constituents, a decrease of
intracellular ATP concentration and a decrease in intracellular
pH due to an increase in the permeability of cell membrane
when
57
Introduction
Review
of Literature
cinnamon oil was applied up to 0.1%. Nonetheless, these

authors did not observe an apparent change on the cell surface
(by electron micrographs) when cinnamon oil was added. In the
same way.
Baskaran et al. (2010) investigated the
antimicrobial effect of low concentrations of transcinnamaldehyde (TC) on Escherichia coli O157:H7 in apple
juice and apple cider. A five-strain mixture of E. coli O157:H7
was inoculated into apple juice or cider at 6.0 log CFU/ml,
followed by the addition of TC (0%v/v, 0.025%v/v, 0.075%v/v
and 0.125%v/v). The inoculated apple juice samples were
incubated at 23 C and 4 C for 21 days, whereas the cider
samples were stored only at 4 C. The pH of apple juice and
cider, and E. coli O157:H7 counts were determined on days 0,
1, 3, 5, 7, 14 and 21. TC was effective in inactivating E. coli
O157:H7 in apple juice and apple cider. At 23 C 0.075 and
0.125 %v/v TC completely inactivated E. coli O157:H7 in apple
juice (negative by enrichment) on days 1 and 3, respectively. At
4 C 0.075 and 0.125 %v/v TC decreased the pathogen counts in
the juice and cider to undetectable levels on days 3 and 5,
respectively. Results indicate that low concentrations of TC
could be used as an effective antimicrobial to inactivate E. coli
O157:H7 in apple juice and apple cider. Abd El-Aziz and Abd
El-Khalek (2010) reported that the cinnamon essential oils
completely inhibited the growth of most yeast and bacteria
strains artificially inoculated in orange juice tested (Candida
albicans, Debaryomyces hansenii, Candida tropicalis,
Saccharomyces cerevisiae, Rhodotorula ruba, Candida
guilliermondii and Staphylococcus aureus). It had moderate
effects against Zygosaccharomyces rouxii, Enterococcus
faecalis and Escherichia coli. They also found that treatment of
orange juice by addition of 4l/ml of orange peel essential oil in
combination with 2.0 kGy gamma irradiation completely
inhibited all the tested microorganisms.
58
Introduction
Materials
and Methods
Materials and Methods

Pomegranate
(Punica
granatum),
tomato
(Solanum
lycopersicum) and carrot (Daucus carota) were chosen for
preparation of fresh fruit juices and fresh vegetable juices.
1. Preparation of Juices
1.1. Preparation of fresh squeezed pomegranate juice
Commercially ripe fresh pomegranates were bought from
a local fruit market at Cairo. Sound fruits were washed with tap
water and the washed fruits were cut-up and the outer leathery
skin, which encloses hundreds of fleshy sacs, was removed. The
arils were manually separated and pressed. The juice was
prepared by a direct squeezing in a commercial juicer (National
juicer blender, Model MJ-130N). The juice samples were
packaged in aluminum foil bags (each 100 ml) and kept frozen
over night, then irradiated.
1.2. Preparation of fresh squeezed tomato juice
Commercially ripe fresh tomatoes were bought from a
local vegetable market at Cairo. All the tomatoes were washed
with running tap water to remove dirt and each was cut into
eight pieces, which were then squeezed by a direct squeezing in
the same commercial juicer. The prepared juice samples were
packaged in aluminum foil bags (each 100 ml) and kept frozen
over night, then irradiated.
59
Introduction
Materials
and Methods
1.3. Preparation of fresh squeezed carrot juice

Commercially ripe fresh carrots were bought from a local
vegetable market at Cairo. The green part on the top of the
carrot and the wounded part were removed and the sound
samples were soaked in tap water for 24 hours. The juice was
prepared by a direct squeezing in the same commercial juicer.
The juice samples were packaged in aluminum foil bags (each
100 ml) and then irradiated
2. Gamma irradiation
Gamma irradiation of the packaged prepared juice
samples was carried out using cobalt 60 irradiator source
(Gamma Chamber 4000 India), located at National Center for
Radiation and Technology ( NCRRT), Nasr City, Cairo, Egypt.
The bags were divided into two groups; the first group was left
without irradiation and considered as control, while the second
were exposed to gamma irradiation as follows:
a) The applied doses for pomegranate juice were 0, 1, 2.0 and
2.5 kGy. The dose rate of the irradiator source at the time of
irradiation was 4.018 kGy /h.
b) Tomato juice bags were irradiated at doses of 0, 1.5, 3.0 and
c) Carrot juice bags were irradiated at doses of 0, 1.5, 3.0 and
Dosimetry was performed using reference alanine dosimeters
traceable to national physical laboratory (NPL), UK
60
Introduction
Materials
and Methods
3. Storage
Both non-irradiated and irradiated samples were
immediately stored in the refrigerator at 4C1 along the period
of study. Periodically, three samples were withdrawn for
microbiological, physiochemical, nutritional and sensory
analysis.
4. Microbiological analysis
The microbiological analysis of pomegranate, tomato and
carrot juice samples including total aerobic bacterial count
(TBC), lactic acid bacteria (LAB), total mold and yeast
(TM&Y),
coliform
bacteria,
Enterococcus
faecalis,
Staphylococcus aureus and Aeromonas hydrophila were
determined during the post irradiation storage period using
standard techniques.
4.1. Total aerobic bacterial counts (TBC)
Total aerobic bacterial counts were determined by
dilution plate method using plate count agar medium according
to APHA (1992). A series of decimal dilutions of original
samples were prepared with sterile saline solution. One ml of
each appropriate dilution was mixed with 15 ml liquid bacterial
count agar that had been cooled to about 45 C, and poured
immediately into sterile 90 mm Petri dishes. After the agar had
hardened, incubation was performed at 30 C for 72 hr. Each
value represents the mean of three samples and results were
expressed as colony forming units (cfu) per milliliter.
4.2. Lactic acid bacteria (LAB)
Lactic acid bacteria (LAB) were counted on Man, Rogosa
and Sharp (MRS) agar medium according to APHA (1992). The
inoculums were plated between two layer of the medium and the
inoculated plates were incubated at 30 C for 24-48 hr. Each of
61
Introduction
Materials
and Methods
the isolates was first tested for catalase by placing a drop of 3%

hydrogen peroxide solution on the cells. Immediate formation of
bubbles indicated the presence of catalase in the cells. Only
those isolates which were catalase-negative were Gramstained.Gram-positive
isolated
colonies
with
typical
characteristics namely pure white, small (2-3 mm diameter) with
entire margins were picked from each plate and transferred to
MRS broth.The cultures were identified according to their
morphological,cultural,
physiological
and
biochemical
characteristics (Kandler and Weiss, 1986).
4.3. Total molds and yeasts
The same procedure of total bacterial count was used for
total moulds and yeasts counts (TM&Y) except that Malt extract
medium containing 100 mg chloramphenicol to suppress
bacterial growth was used. Then the plates were enumerated
after incubation at 25C for 3-5 days APHA (1992). Each value
represents the mean of three samples and results were expressed
as colony forming units (cfu) per milliliter.
4.4. Total Coliform
The total coliform bacteria were determined using
MacConkey broth by Most probable number (MPN) technique
using three tubes according to WHO ( 1993 ). The inoculated
tubes were incubated at 37C for 24-48h. The presence of acid
(yellow color) and gas (in Durhams tubes) indicate positive
results.
4.5. Escherichia coli
E. coli was determined by the MPN technique. For this,
positive MacConkey broth tubes were gently agitated and a
loopful from this culture was transferred to MacConkey Broth
tubes. The tubes were incubated at 44.4C for 48h (APHA
,1992)
62
Introduction
Materials
and Methods
and examined for gas formation and acid production. Positive

cultures were streaked on to Eosine methylene blue agar and
incubated at 37C for 24h. The colonies which were greenish,
metallic, sheen in reflected light and blue black center were
confirmed by IMViC tests according to (APHA, 1992). E.coli
colonies are Indole positive, Methyl red positive, VogesProskauer negative, cannot utilize citrate as a sole carbon source
and can grow at 44.4 C.
a) Indole Production test (tryptophan hydrolysis):
Indole test is a test for the production of indole from
tryptophan . 5ml tryptone water test tubes were inoculated with
E.coli isolates and incubated at 37C for 2-7 days. 0.5ml
Kovacs indole reagent was added to each tube. The tubes were
shaked gently and then allowed to stand. In the presence of
indole, a deep red or pink colour appears in the upper layer only
recorded as indole positive result.
b) Methyl red test:
Five ml glucose phosphate broth tubes were inoculated
with E.coli isolates and incubation at 37C for 2-7 days. Five
drops of the methyl red indicator was added to 5ml of broth
culture, a red colour is described as positive.
c) Voges-Proskauer(VP) test :
Voges-Proskauer test is a test for the production of
acetylmethylcarbinol from glucose. E.coli isolated were
inoculated into 5ml glucose phosphate broth and incubated at
37C for 2-7 days. To 5ml culture, 1ml of Barritts reagent was
added. The tubes were gently shaked, in which any
acetylmethylcarbionl presented became oxidized to diacetyl.
63
Introduction
Materials
and Methods
The diacetyl will combine with arginin, creatine and gave a red
colour which record as a positive result.
d) Simmons citrate utilization test:
Each E.coli isolates was streaked over the surface of a
slant containing Simmons citrate agar medium which used for
citrate utilization as a carbon source, and which contain
bromothymol blue as pH indicator. After inoculation and
incubation at 37C for 2-7days, if the microbe was able to grow
and can utilized a citrate, the medium changes from green to
bright blue.
4.6. Enterococcus faecalis
Enterococcus faecalis was enumerated on Kanamycin
Aesculin Azid Agar (KAAA) medium using surface spreading
technique according to Mossel (1978). The inoculated plates
were incubated at 37 C for 16-24 hr. The porcelaneous
colonies, which are surrounded by black haloes, were counted as
Enterococcus faecalis. Confirmation of E.faecalis was carried
out by catalase test. In this test, 1 ml of fresh hydrogen peroxide
solution was placed in a small clean test tube and 1 ml of each
Enterococcus broth culture was added. Effervescence, caused by
the liberation of free oxygen as gas bubbles, indicates the
presence of catalase in the culture under test. Enterococcus
faecalis also shows chainforming Gram positive cocci cells
and can grow at 45C1.
4.7. Staphylococcus aureus
Staphylococcus aureus was counted on Bairedparker
agar (BPA) medium using surface spreading technique
according to ICMSF (1978). The plates were incubated at 37C
for 24-48 hr. The black colonies surrounded by a clear zone
were counted as Staphylococcus aureus. Confirmation was
carried out by Gram
64
Introduction
Materials
and Methods
staining and by coagulase test using tube method. In this

method, 0.5 ml of rabbit plasma was added to 0.5 ml of 18-24 hr
broth culture in a glass tube.Tubes were incubated at 37C and
visually examined for clotting after 1 hr and at intervals for up
to 24 hr.
4.8. Aeromonas hydrophila
Aeromonas hydrophila was counted on starch ampicillin
agar medium (SAA) using plate surface spreading technique
according to Palumbo et al. (1985). Appropriate dilutions of the
fruit juice were surface plated on SAA and subsequently
incubated at 28C for 24 h. After incubation, the plates were
flooded (5 ml) with Lugol iodine solution and amylase-positive
colonies (those having a clear zone surrounding the colony)
were scored as presumptive A. hydrophila. These colonies are
typically 3 to 5 mm in diameter and yellow to honey colored.
Aeromonas hydrophila colonies was confirmed by their ability
to hydrolyze aesculin broth, voges-proskaure(VP) test,
production of acid from L- arabinose, sucrose and mannitol as
described by Collins et al. ( 1989).
5. Isolation and purification of yeasts
One ml of juice was aseptically added to 9 ml of saline
solution ( 8.5 % Nacl), and serial dilutions was made from
appropriate dilutions ( 10-3 and 10-4 ) one ml was inoculated into
Petri dishes and poured with malt extract agar medium
containing 100 mg chloramphenicol to suppress bacterial
growth. The plates were incubated at 25C for 3-4 days.
Selected colonies (single and not similar) were subcultured on
the surface of the same medium (individually) and incubated at
25C for 3-4 days. The pure culture was streaked on malt extract
agar (containing chloramphenicol) slants and incubated at 25C
for 24 hours, then kept at 4C until identification.
65
Introduction
Materials
and Methods
6. Identification
6.1. Identification of yeast isolates
Yeast isolates were identified according to keys of Barnett
et al. (1990), confirmed and renamed by Barnett et al. (2000)
and Kurtzman and Fell (2000). The identification was
achieved at Food Microbiology Laboratory, NCRRT.
6.2. Incubation temperatures
Yeast are usually incubated at 25C, although optimum
temperature for growth is high for some yeast and lower for
others. Tubes of malt extract broth were inoculated with yeast
isolates and incubated at 25, 30, 35, 37 and 42C for 3-4 days.
6.3. Microscopical examination
Microscopical appearance of non-filamentous vegetative
cells was examined. A young growth culture (onedayold) was
inoculated into 30 ml of yeast extractglucosepeptone medium
in a 100 ml conical flask. The culture was examined
microscopically after incubation for 2-3 days, at 25C.
Microscopical examination for filamentous growth of
ballistoconidia,
chlamydospores,
pseudohyphae
and
blastospores was done after inocubation of a plate containing
corn meal agar with tween80 and incubation at 25C for 3-7
days.
6.4. Biochemical tests
Assessing the ability to use nitrogen compounds for aerobic
growth.
One drop of growth from the yeast carbon base (YCB)
tube was added to a fresh tube of (YCB), the suspension was
used to
66
Introduction
Materials
and Methods
inoculate the nitrogen assimilation medium. Nitrate, nitrite, LLysine and creatine were used as test substates, the nitrogenous
test substrates were added to give a concentration of 5 mM.
Media were adjusted initially to pH 5.5 and incubated at 25C
for 7 days. Growth was determined by visual observation of
turbidity in the broth culture.
Carbohydrate assimilation and fermentation tests
One drop of growth from yeast nitrogen base (YNB)
broth cultures was inoculated into a fresh tube of (YNB) and
mixed thoroughly. The inoculums for the assimilation and
fermentation of carbohydrates was taken from the second
(YNB) broth tube and added to tubes filled with 10 ml of broth
medium and 5 mM of the tested sugar.
Urease test
This test was carried out by inoculating a slant of
Christensens urea agar and incubating the culture at 25C for 7
days to detect change in the color of the medium from yellow to
red.
Cycloheximide sensitivity test
Each isolate was subcultured on malt extract agar
containing 0.01% and 0.1% of cycloheximide and incubated at
25C for 7 days to determine the tolerance of the yeast to
cycloheximide.
7. Determination of D10 - values
The radiation decimal reduction dose (D10- values) for the
identified yeasts were determined from dose response curves
and from the regression linear equation (Lawerence, 1971).
67
Introduction
Materials
and Methods
7.1. Preparation of inoculum

The stock cultures of the identified isolates were
separately activated by growing each in 50 ml malt extract broth
at 25C for 3-5 days. The culture broths were diluted with
sterilized saline to obtain suspension of approximately 107
cfu/ml and used for inoculation of the samples (work culture
suspension). To know the concentration of the work culture
suspension, 1 ml of appropriate dilutions was spreaded onto the
surface of prepared plates of malt extract agar and incubated at
25C for 3-5 days.
7.2. Inoculation
Under aseptic condition, 180 ml of juice (from which the
yeast was isolated) were placed in aluminum foil bags and well
sealed. These bags were exposed to 10 kGy of gamma radiation
to eliminate natural microflora. After irradiation, 20 ml of each
work culture suspension were aseptically added to each juice
bag.
7.3. Irradiation
After inoculation, 10 ml of each inoculated juice samples
were packed in sterilized test tubes. They were exposed to
different irradiation doses (0.0, 0.5, 1.0, 1.5, 2.0 and 2.5 kGy).
Three replicates of each yeast were used in each dose.
7.4. Plating
After irradiation, decimal dilutions were done. 1 ml of
selected dilutions of each yeast under testing was transferred ( in
duplicates ) into Petri dishes and poured by Malt extract agar
medium and incubated at 25C for 3-5 days.
68
Introduction
Materials
and Methods
7.5. D10 value

The D10- values for the isolates and identified yeasts
were determined from dose response curves and from the
regression linear equation (Lawerence, 1971).
Y=abx
D10 value = - 1/b

xy nx y
b =
x
n x
Where:
X = Dose level (kGy)
Y = log number of bacterial surviving after receiving x amount
of radiation
n = number of calculated point
8. Nutritional analysis
8.1. Anthocyanin content in pomegranate juice
The total anthocyanins were estimated by pH differential
method using two buffer systems: potassium chloride buffer, pH
1.0 (25mM) and sodium acetate buffer, pH 4.5 (0.4 M)
according to Giusti and Wrolstad, (2001). The samples were
diluted by a potassium chloride buffer until the absorbance of
the sample at 510 nm wave length was within the linear range of
the spectrophotometer (ATI unicam, 5600 series UV/VIS
spectrophotometer, Model (V-200-RS)). This dilution factor
was later used to dilute the sample with the sodium acetate
buffer. The
69
Introduction
Materials
and Methods
wavelength reading was performed after 15 min of incubation,

diluted in the two different buffers and at two wavelengths of
510 nm and 700 nm. The total anthocyanins content was
calculated as follows:
Total anthocyanins =
Where
A = ( A510 A700 )pH1.0 ( A510 A700 ) pH4.5
MW : molecular weight ( 449.2 )
DF : dilution factor
MA : molar absorptive coefficient of cyaniding-3-glucosid
( 26.900).
Results were expressed as mg cyaniding-3-glucoside 100g-1 of
juice.
8.2. Lycopene Content in Tomato juice
For determination of lycopene content, 4 ml of fresh
tomato juice was put in a 200- ml flask wrapped with aluminum
foil to keep out light. A 100 ml mixture of hexaneacetone
ethanol, 2:1:1 (v/v %) was added to the flask and agitated
continuously for 10 min. After that 15 ml of water was added
followed by another 5 min agitation. The solution was separated
into polar and non polar layers. Non polar layer, i.e. hexane
solution containing lycopene was filtered into by filter paper;
the filtrate was then diluted with a mixture of hexane-acetoneethanol (2:1:1, vol/vol %). Lycopene concentration was
estimated by measuring the absorbance of the hexane solution
containing lycopene at 472 nm on a spectrophotometer (ATI
unicam, 5600 series UV/VIS spectrophotometer, Model (V-200RS)). The
70
Introduction
Materials
and Methods
lycopene was quantified by use of a standard linear curve (R2 =

0.9982) of lycopene solution in hexane in concentrations from
0.25 to 1.25 g/mL. The contents of lycopene were expressed as
milligrams per 100 g wet weight (Markovic et al., 2006).
8.3. Total carotenoids content in carrot juice
10 ml of fresh carrot juice were blended for 2 min with 40
ml of chloroform and methanol (1:3 v/v) mixture for
homogenization .The homogenizate was centrifuged at 4800
rpm (3218 g) for 15 min. The lower layer was withdrawn from
the test tube with a syringe and diluted with chloroform and
methanol (1: 3 v/v). Total carotenoids were determined
spectrophotometricaly at 450 nm using ATI unicam, 5600 series
UV/VIS spectrophotometer, Model (V-200-RS) and the amount
of Carotenoids were calculated with reference to a standard
curve based on beta carotene (Sharon Raber and Kahn,
1983).
8.4. Ascorbic acid measurement
Ascorbic acid was determined using 2, 6 dichlorophenol
indophenols reagent according to the method described by
AOAC (2000). Where, 10 ml of the juice was taken and made
up to 100 ml with 3% oxalic acid, the sample was filtered over
charcoal, 10 ml of the filtrate was taken and then titrated by 2, 6
dichlorophenol indophenols reagent till faint pink color was
observed.
Calculations :
where :
a = Ascorbic acid mg / 100 g or ml
b = Titration
71
Introduction
Materials
and Methods
c = Dye factor
d = Volume made up
e = Aliquot of extract
f = Wt or Volume of sample
9. Physiochemical analysis
9.1. Viscosity
The viscosity of the juice was measured using a
(Brookfield digital Rheometer (Model DV-II, Brookfield
Engineering Laboratories, Inc., Stonghton, MA).
Viscosity measurements were made by using 8-16 ml of juice
samples in small sample adapter. The temperature was adjusted
at 20C. The temperature degree was adjusted using water bath
circulator and coolers. Each sample measured against nonirradiated control.
9.2. pH
Changes in pH value of juices during storage were
determined by a Bench pH meter (pH 211 Microprocessor pH
meter, Hanna instruments) and pH electrode (Epp-1) at room
temperature AOAC (2000).
10. Sensory evaluation
Unirradiated and irradiated juice samples were given to
panelists immediately after irradiation and during subsequent
refrigeration storage for sensory evaluation. The procedure
carried out for this evaluation was similar to that described by
Min et al. (2003). Ten panelists belonging to the Department of
Food Microbiology at the National Center for Radiation
Research
72
Introduction
Materials
and Methods
and Technology Cairo, Egypt were participated in the sensory

tests. Fifteen milliliters of each sample were served into 20 ml
ultra clear polypropylene containers with polyethylene screwcap (Deltalab) coded with three digits randomly numbered;
moreover, a glass containing potable water and a piece of nonsalted cracker were provided to panelists for eliminating the
residual taste between samples. The panelists were asked to rate
the preference of odor, color, taste and overall acceptability in a
hedonic scale from 0 to 9. where ( 9= like extremely, 8= like
very much, 7= like moderately, 6= like slightly, 5= neither like
nor dislike, 4= dislike slightly, 3= dislike moderately, 2= dislike
very much, 1= dislike extremely). A score of 4 or below was
regarded as unacceptable and taken to indicate the end of shelflife.
11. Combination treatments
Fresh carrot juice was selected for the experiment in
which an attempt was made to test whether the cinnamaldehyde
can be applied as natural preservative in combination with
irradiation to improve microbial quality of carrot juice. The
carrot juice was divided into three groups, the first group was
the juice without any treatment and used as negative control
group, the second group prepared in which the concentration of
the cinnamaldehyde was 8 l. 100 ml-l, the third group was
exposed to gamma irradiation at dose levels of 3.0 kGy with the
addition of the cinnamaldehyde (8 l. 100 ml-l). Total aerobic
bacterial count, lactic acid bacterial count and total molds and
yeasts, coliform bacteria, E.coli, Staph. aureus. and Aeromonas
hydrophila were examined according to the previously
mentioned methods.
73
Introduction
Materials
and Methods
12. Statistical analysis

The significance of the data with different factors was
evaluated using Two-way analysis of variance ANOVA. All
analyses were performed with SAS software package version
6.12 (SAS, 1997).
Media and reagents used in microbiological analysis
Media used (the contents of each medium were in g/L)
1. Plate count agar
Yeast extract powder
2.5
Tryptone
5.0
Dextrose
1.0
Agar
15
pH 7.00.2
The ingredients were dissolved in the water by steaming, the pH

was adjusted and the medium was sterilized by autoclaving at
121C for 15 min.
2. Man, Rogosa and Sharp (MRS)
Peptone
10.0
Lab lemco powder
8.0
Yeast extract
4.0
D-Glucose
20.0
Tween 80
1ml
74
Introduction
Materials
and Methods
Dipotassium hydrogen phosphate
2.0
Sodium acetate
5.0
Triammonium citrate
2.0
Magnesium sulphate , hydrated (MgSO4.7H2O)
0.2
Manganese sulphate , hydrated (MgSO4.7H2O)
0.05
Agar15.0
pH 6.00.1
121C for 15 min.
3. Malt Extract Agar
Malt extract
30
Mycological peptone
Agar
15
pH 5.40.2

121C for 15 min. After cooling the medium to 48C and
before pouring. 100 mg of chloramphenicol were added to
suppress bacterial growth.
4. MacConkeys broth
Peptone
20.0
Bile salts
5.0
75
Introduction
Materials
and Methods
Sodium chloride
5.0
Lactose
10.0
Bromocresol purple
0.01
pH 7.10.1
The ingredients were dissolved ( except indicator ) in the water

by steaming , the pH was adjusted , the indicator was added.
The medium was mixed well and distributed (7-9) ml in test
tubes each containing Durhams tube, then sterilized by
autoclaving at 121C for 15 min.
5. Eosin Methylene blue agar
Peptone
10.0
Lactose
10.0
2.0
Eosin yellowish
0.4
Methylene blue
0.065
Agar
15.0
pH 6.8 0.2

was adjusted , and the medium was sterilized by autoclaving at
121C for 15 min. The medium was cooled to 50C and shacked
in order to oxidized the methelene blue (restore its blue color)
and suspended the precipitate which is an essential part of the
medium and poured in plates.
76
Introduction
Materials
and Methods
6. Tryptone water broth

Tryptone
10.0
Sodium chloride
5.0
pH 7.20.2

was adjusted. The medium was distributed (7-9) ml in test tubes,
then sterilized by autoclaving at 121C for 15 min.
7. Glucose phosphate broth
Peptone
5.0
D-Glucose
5.0
5.0
pH 7.50.2
was adjusted. The medium was distributed (7-9) ml in test tubes,
then sterilized by autoclaving at 121C for 15 min.
8. Simmons citrate agar
Sodium chloride
5.0
Magnesium sulphate
0.2
Ammonium dihydrogen phosphate
0.2
Sodium citrate
2.0
Bromothymol blue
0.08
77
Introduction
Materials
and Methods
Sodium ammonium phosphate
0.8
Agar
15.0
pH 7.00.2
The ingredients were dissolved ( except indicator ) in the water

by steaming, the pH was adjusted, the indicator was added. The
medium was mixed well and distributed in test tubes, then
sterilized by autoclaving at 121C for 15 min and allowed to set
in a slop position
9. Kanamycine aesculine azide agar
Tryptone
20.0
Yeast extract
5.0
Sodiumchloride
5.0
Sodium citrate
1.0
Aesculin
1.0
Ferric ammonium citrate
0.5
Kanamycin sulphate
0.02
Sodium azide
0.15
Agar
15.0
pH 7.10.2

121C for 15 min. The medium was cooled at 48C and poured
in plates.
78
Introduction
Materials
and Methods
10. Baird parkers medium (Egg-yolk tellurite glycine agar)

Tryptone
10.0
Lab-lemco meat extract
5.0
Yeast extract
1.0
Lithium chloride
5.0
Glycine
12
Agar
20.0
pH 6.80.2

121C for 15 min. The medium was allowed to cool to 48-50C
and aseptically the following filtered sterilized solutions were
added ( to each 90 ml of the basal medium )
Sodiumpyruvate 20% (w/v)
5.0ml
Potassium tellurite 1% ( w/v)
1.0ml
Egg-yolk emulsion
5.0ml
The complete medium was well mixed and poured immediately

in plates
11. Starch ampicillin agar (SAA)
Beef extract
1.0
Protease peptone
10.0
Sodium chloride
5.0
79
Introduction
Materials
and Methods
Phenol red
0.025
Soluble starch
10.0
Agar
15.0
pH 7.40.1

121C for 15 min. The medium was cooled at 48C, ampicillin
was added to achieve a concentration of 10mg/L.
12.Aesculine hydrolysis broth
Peptone
10.0
Sodium citrate
1.0
Aesculine
1.0
Ferric citrate
0.05
pH 7.00.1
The ingredients were dissolved in the water by steaming; the pH

was adjusted and distributed (7-9) ml in test tubes, then was
sterilized by autoclaving at 121C for 15 min.
13. Christensens urea agar (urea agar base)
Peptone
1.0
Dextrose
1.0
Sodium chloride
5.0
Disodium phosphate
1.2
80
Introduction
Materials
and Methods
Potassium dihydrogen phosphate
0.8
Phenol red
0.012
Agar
15.0
pH 6.8
Urea solution 40% (filter sterile)
81
5ml
Introduction
Materials
and Methods
Reagents used
1-kovacs indole reagent
Di methyl amino benzaldehyde
5g
Pentanol ( amyl alcohol )
75ml
Conc. HCL
25ml
The aldehyde was dissolved in alcohol, then slowly acid was

added, then store in refrigerator.
2- Methyl red reagent
Methyl red
0.1g
Ethanol (95%)
300ml
Distilled water
500ml
The methyl red was dissolved in the ethanol and maked up to

500 ml with distilled water.
3- Voges-proskaure test ( Barritts ) reagent
Potassium hydroxide 16% solution
- naphthol 6% in 95% ethanol
4- Catalase test reagent
H2O2 (3% aqueous solution 10 vol.)
5-Lugols iodine reagent
Iodine
1g
Potassium iodide
2g
82
Introduction
Materials
and Methods
Distilled water
300 ml
The aldehyde was dissolved in alcohol, then slowly acid was

added, then store in refrigerator.
Lycopene standard
Were obtained from Sigma Chemical Co. ( St. Louis,MO ).
-carotene standard
Were purchased from Fluka (Deisenhofen,Germany).
2,6 dichlorophenolindophenol
Were purchased from Fluka (Deisenhofen,Germany).
Cinnamaldehyde
The Cinnamaldehyde used in this study were obtained from
Himedia laboratories pvt.Ltd. (23,Vadhani Ind.Est.,LBs Marg ,
Mumbai-400086, India )
83
Introduction
Results
and Discussion
Results and Discussion

1. Isolation and Identification of yeasts
Natural vegetable juices are highly perishable and have a
limited shelf life, rarely exceeds one day even under a cold
chain system. Deterioration in the quality and spoilage of these
vegetable juices is mainly related to the growth of the acidloving or acid-tolerant bacteria and fungi (yeasts and moulds).
However, Tournas et al. (2006) reported that the most
microorganisms found in the various types of juices were yeasts.
Yeast spoilage of fruit juice can result in formation of haze,
production of CO2 and off-odors, and changes in color
(Grinbaum et al., 1994).
Therefore, an attempt have made in the present study to
isolate and identify the yeast flora from the selected fresh juices,
i.e. pomegranate, tomato and carrot juices.
Twenty yeast colonies were isolated from the juices under
investigation, pomegranate juice (7 colonies), tomato juice (8
colonies) and carrot juice (5 colonies). They were identified
according to their morphological characteristics, microscopical
appearance and biochemical tests (table 3). The results (table 4)
and figures (6, 7, 8, 9, and 10) of the identification revealed that
the 7 colonies from pomegranate juices were Candida tropicalis
(4 colonies) and Debaryomyces hansenii (3 colonies).
Meanwhile, the 8 colonies from tomato juices were
Saccharomyces cerevisiae (5 colonies) and Trichosporon
pullulans (3 colonies). While, the 5 colonies from carrot juice
were Rhodotorula glutinis. Many other investigators isolated
several yeasts from fruit and vegetable juices.
84
Introduction
Results
and Discussion
Beech and Davenport (1970) isolated three new Candida

species (C. anglica, C. cidri and C. pomicola) from apple cider.
Mendoza et al. (1982) stated that Rhodotorula, Pichia, Candida
and Saccharomyces were frequently isolated from pasteurized
fruit juices. Warnasuriya et al. (1985) isolated Saccharomyces
cerevisiae from tomato sause. Candida and Saccharomyces spp.
have often been reported as spoilage-causing organisms in citrus
juices (Parish and Higgins, 1989; Teller and Parish, 1992).
Wade et al. (2003) isolated Debaryomyces hansenii and
Trichosporon pullulans from tomatoes.Tournas et al. (2006)
reported that the majority of fruit salad samples (97%) were
contaminated with yeasts at levels ranging from <2.0 to 9.72
log10 of cfu/g. Frequently encountered yeasts were Pichia spp.,
Candida pulcherrima, C. lambica, C. sake, Rhodotorula spp.,
and Debaryomyces polymorphus and yeasts commonly found in
fruit juices were C. lambica, C. sake, and Rhodotorula rubra.
Other investigators have also reported the yeast spoilage of fruit
juices.
85
Introduction
Results
and Discussion
Table (3): Biochemical characteristics of the isolated yeasts:

1
+
+
+
+
+
+
-
+
+
V
V
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
Yeast isolate
0.1%
Cycolohexmide
growth
0.01%
Cycolohexmide
growth
Lysine growth
Nitrite growth
Nitrate growth
Manitol growth
Starch growth
Raffinose
growth
Lactose growth
Sucrose growth
Maltose growth
D-Galactose
growth
D-Glucose
growth
Starch
fermentation
Raffinose
fermentation
Lactose
fermentation
Sucrose
fermentation
Maltose
fermentation
D-Galactose
fermentation
D-Glucose
fermentation
Pseudohyphae
Vegetative cell
Growth at 42C
Growth at 37C
Growth at 35C
Growth at 30C
Growth at 25C
Urea hydrolysis
Budding cell
Budding cell
Budding cell
Budding cell
Budding cell
V
+
+
+
+
-
+
+
+
+
+
-
+
+
+
+
+
+
V
V
V
+
+
+
+
+
-
V= very weak
86
Introduction
Results
and Discussion
Table (4): Identification of yeasts isolated from juices

Type of juice
isolate
Pomegranate
juice
Tomato juice
2
5
1
4
3
Carrot juice
No. of
isolates
4
3
5
3
5
Isolated yeast
Candida tropicalis
Debaryomyces hansenii
Saccharomyces cerevisiae
Trichosporon pullulans
Rhodotorula glutinis
Microscopic examinations
Photo(1):Candida tropicalis
87
Introduction
Results
and Discussion
Photo (2):Debaryomyces hansenii
Photo (3): Saccharomyces cerevisiae
88
Introduction
Results
and Discussion
Photo (4): Trichosporon pullulans
Photo(5):Rhodotorula glutinis
89
Introduction
Results
and Discussion
2. Radiation decimal reduction dose ( D10- value)

Microorganisms differ greatly in their resistance to
radiation. The radiation-decimal reduction dose (D10-value) of
microorganisms is defined as the radiation dose (kGy) required
to reduce the population of that microorganism by a 10 fold, or
to kill 90% of the total number of that microorganism. It was
found that the radiation resistance of yeasts higher than that of
molds and bacterial pathogens. Thus, most research efforts
related to irradiation of juices have targeted spoilage organisms
such as yeasts rather than bacterial pathogens (Grinbaum et al.,
1994).
In this work, identified yeast isolates (Rhodotorula
glutinis, Candida tropicalis, Trichosporon pullulans,
Debaryomyces hansenii and Saccharomyces cerevisiae) isolated
from pomegranate, tomato and carrot juices were studied for
their resistance to gamma radiation. The so-called dose-response
curve of each yeast was determined in saline solution and in the
juice where it was isolated from. It was constructed by plating
the number of surviving cells against radiation doses used (0.0,
0.5, 1.0, 1.5, 2.0 and 2.5 kGy). The D10-value of each yeast was
determined from own dose-response curve.
The effect of increment doses of gamma irradiation (0.0,
0.5, 1.0, 1.5, 2.0 and 2.5 kGy ) on the viable count of
Rhodotorula
glutinis,
Candida
tropicalis,Trichosporon
pullulans,
Debaryomyces hansenii and Saccharomyces
cerevisiae are tabulated in tables 5, 6, 7, 8, 9 and illustrated in
figures 11, 12, 13, 14, 15, respectively. It is clear that, generally,
log viable counts of tested microorganisms decreased with
increasing radiation dose but at different levels indicating that
these yeasts greatly differ in their sensitivity to gamma
irradiation. From the data it was found that the D10-values (
calculated from linear
90
Introduction
Results
and Discussion
regressions and equations ) of Rhodotorula glutinis, Candida

tropicalis, Trichosporon pullulans, Debaryomyces hansenii and
Saccharomyces cerevisiae in saline solution were 0.92, 0.84,
1.25, 1.29 and 1.01 kGy, respectively. Meanwhile the D10values of these yeasts in the juices were 1.34, 0.98, 1.40, 1.63
and 1.34 kGy, respectively. It is obvious that Debaryomyces
hansenii had the highest D10-values, followed by Trichosporon
pullulans indicating that they had the highest radiation
resistance in comparison with the other studied yeasts. While,
Candida tropicalis had the lowest radiation resistance, i.e. had
the highest sensitivity to radiation. Ingram and Roberts (1980)
mentioned that there are differences in radiation resistance
among microorganisms, i.e. from genus to another and from
species to another of the same genera.
91
Introduction
Results
and Discussion
Table (5): Effect of incremental irradiation doses on the viable count

(cfu.ml-1) of Rhodotorula glutinis in carrot juice
Irradiation
doses (kGy )
Broth
juice
cfu.ml-1
Log N
cfu.ml-
0.0
4.9 x106
6.69
5.2 x105
5.71
0.5
2.3 x106
6.36
4.7 x105
5.67
1.0
5.7 x105
5.75
1.1x105
5.06
1.5
2.7 x105
5.43
5.3 x104
4.72
2.0
4.1 x104
4.61
3.1 x104
4.49
2.5
1.0 x104
4.02
7.8 x103
3.89
D10
D10=0.92
Log N
D10=1.34
Fig. (6): Effect of incremental irradiation doses on the viable count

( cfu.ml-1) of Rhodotorula glutinis in carrot juice.
92
Introduction
Results
and Discussion

(cfu.ml-1 ) of Candida tropicalis in pomegranate juice
Irradiation
doses
(kGy )
Broth
cfu.ml-1
0.0
juice
Log N
cfu.ml-
2.3 x107
7.36
2.6x106
6.41
0.5
1.4 x107
7.16
2.2 x106
6.34
1.0
9.9x105
5.99
1.0x105
5.00
1.5
6.6 x105
5.81
7.5 x104
4.87
2.0
7.7 x104
4.88
1.7 x104
4.23
2.5
4.4 x104
4.64
1.4 x104
4.14
D10
D10=0.84
Log N
D10=0.98
Fig. (7 ): Effect of incremental irradiation doses on the viable count

(cfu.ml-1 ) of Candida tropicalis in pomegranate juice
93
Introduction
Results
and Discussion

(cfu.ml-1 ) of Trichosporon pullulans in tomato juice
Irradiation
doses (kGy )
Broth
juice
cfu.ml-1
Log N
cfu.ml-
0.0
6.9x106
6.83
4.5x106
6.65
0.5
3.4 x106
6.53
1.9 x106
6.27
1.0
1.1x106
6.06
9.0x105
5.95
1.5
8.7 x105
5.93
2.0x105
5.30
2.0
1.2 x105
5.09
1.8x105
5.25
2.5
8.5 x104
4.92
8.0x104
4.90
D10
D10=1.25
Log N
D10=1.40

(cfu.ml-1 ) of Trichosporon pullulans in tomato juice.
94
Introduction
Results
and Discussion

( cfu.ml-1 ) of Debaryomyces hansenii in pomegranate juice
Irradiation
doses
(kGy )
Broth
juice
cfu.ml-1
Log N
cfu.ml-1
Log N
0.0
1.0x106
6.00
5.0x105
5.69
0.5
6.0 x105
5.77
3.0 x105
5.47
1.0
7.1x104
4.85
2.8x105
5.44
1.5
5.5x104
4.74
6.0 x104
4.77
2.0
3.3x104
4.51
2.4 x104
4.38
2.5
1.2x104
4.07
2.2x104
4.34
D10
D10=1.29
D10=1.63

( cfu.ml-1 ) of Debaryomyces hansenii in pomegranate juice.
95
Introduction
Results
and Discussion
Table (9): Effect of incremental irradiation doses on the viable

count
(cfu.ml-1 ) of Saccharomyces cerevisiae in tomato
Irradiation
doses
(kGy )
Broth
juice
cfu.ml-1
Log N
cfu.ml-
Log N
0.0
2.7x107
7.43
9.0x106
6.95
0.5
1.2x107
7.07
4.0x106
6.60
1.0
2.0x106
6.30
1.4x106
6.14
1.5
1.3x106
6.11
1.3x106
6.13
2.0
1.7x105
5.23
1.7x105
5.23
2.5
1.3x105
5.11
1.5x105
5.17
D10
D10=1.01
D10=1.34
juice
Fig. (10): Effect of incremental irradiation doses on the viable

count ( cfu.ml-1 ) of Saccharomyces cerevisiae in tomato juice.
96
Introduction
Results
and Discussion
These results indicated that the D10-values of the yeast

under investigation in juices were higher than its values in broth
indicating that the juices caused some of protection to the yeasts
against radiation as a general trend. Also, the juice as a
suspending medium may contain certain compounds that acted
as protective agents or scavengers which gave protection against
radiation damage of the yeasts cells. Niven (1985) concluded
that microbial cells were generally radiation resistant in
complex media than in simple systems. Similar, the obtained
D10-values have been found by many investigators. Vlad et al.
(1971) found that D10-value for saccharomyces sp. was 1.4 kGy.
Abd El-Aziz (2002) found that D10-value of Saccharomyces
cerevisiae in saline solution was 1.85 kGy, and Candida
tropicalis was 1.31 kGy. Youssef et al. (2002) reported that D10value of Candida tropicalis in saline was 1.23 kGy , where in
mango pulp was 1.37 kGy and that of Rhodotorula glutinis was
1.76 kGy in saline while 2.15 kGy in mango pulp.
Various environmental factors may affect the sensitivity
of microorganisms to radiation among them the medium used
for inoculation, radiation, temperature, condition of irradiation
(presence or absence of oxygen), etc. The difference of the D10values attributable to media may, at a very basic level, also be
attributable to the availability of water (aw) in the medium.
Generally, at a higher water activity, microorganisms are more
sensitive to radiation because of higher presence of free radicals.
Also, in complex food systems, the chemicals affect the
sensitivity of microorganisms to radiation. For example,
proteins in the food acted as quenchers of the radicals formed by
irradiation, leaving fewer ions to react with organisms (Urbain,
1986). Farkas (1990) reported that the response of a microbial
cell and hence its resistance to ionizing radiation depends on:
97
Introduction
Results
and Discussion
1) The nature and amount of direct damage. 2) The number,

nature and longevity of radiation induced reactive chemical
species and the inherent ability of the tolerate or accurately
repair either of the above. 3) The influence of intra-and extracellular environment of any of the above.
Determination of the D10- value for microorganisms from
survival curves (dose-response curve) is very important and
useful in:
1- Calculating the lethal and sub-lethal dose of microorganisms.
2- Providing information about the relative resistance of
particular microorganisms to ionizing radiation.
3- Calculating the exact radiation dose required for 5-log cycles
reduction of pathogenic bacteria, mycotoxin production fungi
and spoilage yeasts, which is very important for the practical
application of ionizing radiation. By knowing the D10-value of
any microorganism and its level in a product (food, feed,
medicinal or pharmaceutical products) one can calculate the
exact irradiation dose required to eliminate that microorganism
from the product.
98
Introduction
Results
and Discussion
3. Irradiation of fresh pomegranate juice

Fresh squeeze pomegranate juice was exposed to different
irradiation doses (1.0, 2.0 and 2.5 kGy). Unirradiated and
irradiated juice samples were stored under refrigeration
condition (4C 1).
3.1. Effect of irradiation and refrigeration storage on the
microbial load of pomegranate juice
Microbial, nutritional, physicochemical and sensorial
properties of these samples were followed during refrigeration
storage. The changes in the microbial load (aerobic mesophillic
bacteria, lactic acid bacteria, yeasts and molds) of pomegranate
juice are shown in table (10). The total log counts of these
microorganisms were 4.90, 4.10 and 3.96 cfu / ml, respectively.
The obtained results were higher than those reported by
Alighourchi et al. (2008 ) who found that the initial mean
populations of total bacterial counts and mold and yeasts counts
for two varieties of pomegranate fresh juice were 3.67, 3.31 and
3.53, 3.00 log cfu/ml, respectively. This is because of the
difference in pomegranate variety, cultivar, climate,..etc.
The effects of gamma irradiation treatment (1, 2 and 2.5
kGy) on the total mesophillic bacteria, lactic acid bacteria, mold
and yeast populations in pomegranate juice are also shown in
table (10). In general, a progressive increase in the dose levels
showed concomitant decrease in the all microbial counts of
pomegranate juice. Irradiation dose of 1 kGy reduced the
number of the initial aerobic mesophillic bacteria by almost 1
log cycle. As well as, it could reduce the lactic acid bacteria by
about 0.16 log cycle.
99
Introduction
Results
and Discussion
Table (10): Effect of -irradiation and subsequent storage (4C1 ) on

the microbial counts ( log cfu/ml ) contaminating
pomegranate juice
Microorganisms
Total
aerobic
mesophilic
bacteria
Lactic acid
bacteria
Total mold
and yeasts
Storage
Irradiation doses ( kGy )
( days )
0.0
1.0
2.0
2.5
4.90aa0.02
3.95ba0.05
2.57ca0.01
<10
5.71ab0.02
4.25bb0.01
2.89cb0.02
<10
14
4.53bc0.06
3.04cc0.01
<10
21
3.14cd0.005
<10
4.10aa0.02
3.94ba0.02
2.04ca0.01
<10
3.98ab0.01
3.90bb0.008
<10
<10
14
3.89 c0.005
<10
<10
21
<10
<10
3.96aa0.05
3.02ba0.01
2.95ca0.02
2.15da0.02
5.22ab.0.0
4.90bb0.005
2.97ca0.01
<10
14
5.05bc0.005
4.79cb0.03
<10
4.90cc0.003
<10
21
R = Samples sensorially rejected .

<10 = below detectable limit of (<10 cfu/ml).
Mean values followed by different superscript (within rows) and different
subscripts (within columns) are significantly different (p < 0.05)
100
Introduction
Results
and Discussion
Also the results indicated that a dose of 2.5 kGy could be

significantly to reduce the total aerobic mesophillic bacteria and
lactic acid bacteria. It reduced the number of the mold and
yeasts by almost 1.81 log cycles. Alighourchi et al. (2008)
reported that 0.5 kGy reduced the total bacterial count of two
pomegranate varieties by almost 0.75 log cycles and reduced the
total mold and yeasts by 0.23 and 0.51 log cycles in the two
pomegranate varieties. They also found that the microbial
population reduced to below the detectable limits at 3.5 kGy,
in all studied juices. Lee et al. (2009) reported that the initial
populations of the total bacterial counts in ready- to- use
tamarind juice were significantly reduced by gamma irradiation
at 1 kGy or above. These results showed that, the inactivation
microorganisms by irradiation in different juices depend on their
compositions.
Table ( 10 ) revealed that the storage of pomegranate
juice at 4C1, showed progressive increase in the viable counts
of aerobic mesophillic bacteria, mold and yeasts in the non
irradiated pomegranate juice reaching 5.71, 5.22 log cfu/ml,
respectively after 7 days. However, these microorganisms
reached almost similar counts in pomegranate juice receiving 1
and 2 kGy but after 14 and 21 days, respectively. Where mold
and yeasts receiving 2.5 kGy reduced to below the detectable
counts (<10) throughout the storage period and this may be due
to the antimicrobial effect of pomegranate juice. Alighourchi et
al. (2008) found that irradiation at 0.5 and 2.0 kGy reduced the
growth rate of bacteria and fungi of the selected pomegranate
juices during the first 3 days of storage at 4C1. Based on the
results of Aziz and Moussa (2002) and kim et al. (2007) the 35 kGy radiation doses may prevent microbial growth in the kale
juice during storage period at 10C. Meanwhile, lactic acid
bacteria decreased during storage. It was considered that the
101
Introduction
Results
and Discussion
decreasing of these bacteria was due to the post-irradiation

effect where the surviving cells that had been damaged by an
irradiation were gradually inactivated, thus not adapting to the
surrounding environment during storage. Byun et al. (2001)
reported that the sublethal damage to cells caused by irradiation
is likely to increase their sensitivity to environmental stress
factor.
pathogenic bacteria contaminating pomegranate juice
The results in table (11) indicated that the most probable
number of coliform bacteria in pomegranate juice was 75
MPN/ml. On the other hand, Escherichia coli, Enterococcus
faecalis, Staphylococcus aureus and Aeromonas hydrophila
were below detectable level in unirradiated (control)
pomegranate juice, and this may be due to the acidic pH. Foley
et al. (2002) found that the counts of Listeria monocytogenes
and Salmonella had decreased by more than 3 to 4 logs at pH
3.6. Also may be due to antibacterial and synergistic action of
the pomegranate constituents. Many investigators all over the
world, found that pomegranate has antimicrobial, antibacterial,
antifungal and antimutagenic properties as well as it is rich in
tannins which have remarkable antimicrobial activity (Machado
et al., 2003 and Voravuthikunchai et al., 2004). Kumar and
Vaithiyanathan (1990) reported that the protein binding
capacity of phenolics as (anthocyanin in pomegranate) caused
inhibition of microbial growth and antimicrobial property.
102
Introduction
Results
and Discussion
Table (11): Effect of -irradiation and subsequent storage (4C1) on

the some pathogens contaminating pomegranate juice
Micro- organisms
Coliforms
( MPN/ml)
Escherichia coli
( MPN/ml )
Enterococcus
faecalis
( cfu / ml )
Staphylococcus
aureus
( cfu / ml )
Aeromonas
hydrophila
( cfu / ml )
Storage
( days )
0
7
14
21
0.0
75
93
R
R

1.0
2.0
<3
<3
<3
<3
<3
<3
<3
R
<3
0
7
14
21
<3
<3
R
R
<3
<3
<3
R
<3
<3
<3
<3
<3
<3
<3
<3
0
7
14
21
<100
<100
R
R
<100
<100
<100
R
<100
<100
<100
<100
<100
<100
<100
<100
0
7
14
21
<100
<100
R
R
<100
<100
<100
R
<100
<100
<100
<100
<100
<100
<100
<100
0
7
14
21
<100
<100
R
R
<100
<100
<100
R
<100
<100
<100
<100
<100
<100
<100
<100
R = Samples sensorially rejected .

< 3 = No positive tubes have been shown in the first three dilutions
<100 = Below detectable level
103
2.5
<3
<3
<3
<3
<3
Introduction
Results
and Discussion
The results (Table 11) also showed that coliform bacteria

present in unirradiated pomegranate juice were eliminated by
the lowest irradiation dose used (i.e.1 kGy) since the MPN/ml
was below the detectable level (< 3) indicating the sensitivity of
coliforms to gamma irradiation. The sensitivity of colifrom
bacteria to gamma radiation has shown by many investigators
(Patterson, 1988; Kamat et al., 2003; Goularte et al., 2004;
Lu et al., 2005 and Abu El Nour, 2007).
During storage of pomegranate juice at 4C1 coliform
bacteria increased to reach 93 MPN/ml at day 7. On the other
hand, the MPN of coliform and all tested pathogens in irradiated
samples were below the detectable level throughout the
refrigerated storage period.
anthocyanin content of pomegranate juice
Anthocyanins are a member of phenolics compounds that
contributes to the red, blue or purple color of many fruits,
including pomegranate juice, and they are well-known for their
antioxidant and antimicrobial activity.
The change in the anthocyanin content of pomegranate
juice as affected by irradiation is tabulated in table (12) and
illustrated in figure (16). The total anthocyanin content of fresh
pomegranate juice (unirradiated control) samples was 28.85 mg
/100 g. This value is within the range of pomegranate juice
anthocyanin content (8.1-36.9 mg/100g) reported by Cam et al.
(2009). Also, Tehranifar et al. (2010) found that the
anthocyanin content of pomegranate juice ranged from 5.5630.11 mg /100g depending on cultivars. The results in the same
table and figure indicated that gamma irradiation at 1.0, 2.0 and
2.5 kGy
104
Introduction
Results
and Discussion
decreased anthocyanin content of pomegranate juice by 7.07,

14.00 and 15.45%, respectively. This decrease could be
attributed to the degradation of anthocyanin compounds by
irradiation. The anthocyanin molecule has many highly reactive
sites for free radical damaging, for example, the phenolic
hydroxyls and the aromatic ring itself. Gamma radiation induced
electron delocalization is shared by the whole system, in this
way; energy is not concentrated in any particular site of the
molecule (Spinks and woods, 1990). During radiation
treatment, a certain number of molecules get modified and this
reflects in products colour reduction. Also, irradiation effects on
anthocyanin pigments depend upon the nature of anthocyanin
for example; diglycosides are relatively stable towards
irradiation dose compared to monoglycosides (Alighourchi et
al., 2008).
Table (12): Total anthocyanin content in pomegranate juice during
storage at 4C1
Storage
( days )
Irradiation dose ( kGy )
0
28.85aa1.27
1.0
26.81ba0.43
2.0
24.81ca0.23
2.5
24.39ca1.17
26.09ab1.09
25.82bb1.63
24.04ca2.03
23.64db0.24
14
22.47bc2.04
21.88cb3.54
19.63dc1.67
21
17.76cc2.80
16.96dd1.48
R = Samples sensorially rejected.

subscripts (within columns) are significantly different (p < 0.05).
105
Introduction
Results
and Discussion
Fig. (11): Total anthocyanin content in pomegranate juice during storage at

4C1.
The results (Table 12) also shows that storage for 7 days
at 4C1 resulted in a significant decrease in the total
anthocyanin content of both the non irradiated and irradiated
pomegranate juice sample, where the loss reached 9.56, 3.69,
3.10 and 3.07 for samples irradiated at 0.0, 1, 2 and 2.5 kGy,
respectively.
Anthocyanin degradation by gamma radiation is widely
documented in bibliography, for example, Thomas (1986)
reported 20% anthocyanin destruction in 2.5 kGy - irradiated
strawberries. Alighourchi et al. (2008) found that gamma
irradiation at all applied doses (0-10 kGy) significantly reduced
total and individual anthocyanins of the six varieties of
pomegranate juices. They added that pomegranate juices that
irradiated at 0.5 kGy and 10 kGy retained approximately 80 and
10% of the anthocyanin content compared with the control,
respectively. In contrast, Ayed et al. (1999) investigated the
effect of irradiation (0-9 kGy) of grape pomace and they
observed that anthocyanin contents increased at all doses of
irradiation,
106
Introduction
Results
and Discussion
especially at 6 kGy. This increase may be due to the extraction

of bound pigments by degradation of the cell wall. Many
investigators indicated that the stability of anthocyanins against
the irradiation was related to juice compositon. The differences
among juices originated from fruits variety, and the relative
stability of an anthocyanin depends on its matrix, structural
features and the processing conditions (Talcott et al., 2003;
Torskangerpoll and Andersen, 2005).
3.4. Effect of irradiation and refrigeration storage on some
physiochemical properties of pomegranate juice
a) Viscosity
The consistency of juices expressed in viscosity is
important for the quality of juices. Table (13) and figure (17)
show the effect of irradiation on the viscosity of pomegranate
juice. The results indicated that pomegranate juice has lower
viscosity 4 cPs( centipoise). This may be because pomegranate
juices do not contain pectin. It is also clear that irradiation at 1
kGy had no effect on the viscosity of pomegranate juice.
Meanwhile, irradiation at 2.0 and 2.5 kGy significantly reduced
the viscosity. Gagnon et al. (1968) showed that irradiation
lower the viscosity of apple juice and therefore increases rate of
filtration in it. Swailam (1998) detected a reduction in the
consistency index of the irradiated mango pulp, and he reported
that it may be taken in consideration for the detection of
irradiated mango pulp at zero time and during storage.
The results ( Table 13 and figure 17) also show that the storage
for 7 days at 4C1 resulted in insignificant decrease in
viscosity of pomegranate juice of both the non irradiated and
irradiated, but
107
Introduction
Results
and Discussion
the viscosity gradually decreased during further storage in all

irradiated samples.
Table (13):
Storage
( days )
Viscosity (cPs) of pomegranate juices treated by

0

1.0
2.0
2.5
4.0aa0.005
4.0aa0.02
3.0ba1.0
2.0ca0.01
4.0aa0.005
4.0aa0.01
3.0ba1.0
2.0ca0.005
14
2.0bb0.01
2.0bb0.01
2.0ba0.01
R
R
2.0cb0.02
2.0ca0.02
21
R = Samples sensorially rejected,
subscripts (within columns) are significantly different ( p < 0.05 ).
Fig. (12): Viscosity (cPs) of pomegranate juices treated by

irradiation during storage at 4C1.
108
Introduction
Results
and Discussion
b) pH
The results of pH values of pomegranate juice are present
in table (14) and figure (18). It is clear from these results that
the mean pH value of fresh pomegranate juice (unirradiated
control) was 3.8 indicating acidic medium. This value is within
the normal range (2.75 4.14) reported for mature pomegranate
as reported by Al. Maiman and Dilshad (2002). Irradiation at
different doses used (1.0, 2.0 and 2.5 kGy) had no effect on the
pH values of the pomegranate juices. Noci et al. (2008) also
found no differences in the pomegranate juices pH and soluble
solids content of the UV irradiated apple juice.
During refrigeration storage, there was significant
increase in the pH values of non irradiated pomegranate juice
samples and those irradiated at 1.0 and 2.0 kGy. This may be
due to the decrease of lactic acid bacteria count during storage.
(Table 14), indicating a relationship between lactic acid bacteria
and pH value (acidity). Yoon et al. (2004) found that the lactic
acid cultures reduced the pH to 4.1 or below and increased the
acidity to 0.65% or higher in tomato juice.
109
Introduction
Results
and Discussion
Table (14): pH value of irradiated and non-irradiated pomegranate

Storage
( days )
0
3.80aa0.005
1.0
3.81aa0.005
2.0
3.82aa0.005
2.5
3.81aa0.005
4.00ab0.01
4.05ab0.025
4.05ab0.015
3.90aa0.005
14
4.01bb0.005
4.02bb0.02
3.90ba0.01
21
4.40cc0.005
3.90da0.02

subscripts (within columns) are significantly different ( p < 0.05 )
Fig. (13): pH value of irradiated and non-irradiated pomegranate

juices during storage at 4C1.
110
Introduction
Results
and Discussion

sensory evaluation of pomegranate juice
Sensory evaluation of irradiated and non irradiated
pomegranate juice was achieved immediately after irradiation
and during subsequent refrigeration storage at 4C1. Panelists
sensory scores are tabulated in table (15) and illustrated in
figure (19). It is clear from these results that irradiation at the
lowest irradiation dose used, i.e. 1.0 kGy almost had no effect
on sensory properties (color, odor, taste and overall
acceptability) of pomegranate juice, indicating that immediately
after irradiation at all irradiation dose there was no significant
difference between non-irradiated samples and those receiving
1.0 kGy. Meanwhile, irradiation at 2.0 and 2.5 kGy
significantly affected sensory characteristics of pomegranate
juice samples as the panelists gave these samples lower scores
in comparison with non irradiated (control) samples, however,
still like very much. Chachin and Ogato (1969) treated grape,
apple and orange juice with different irradiation doses of gamma
radiation. They found browning in apple juice after 5 kGy and
evident loss of grape juice anthocyanin and orange juice betacarotene after 10 kGy. Miller and Mc Donald (1996) reported
that the juice from irradiated grape fruit was acceptable after 3.0
kGy but declined in quality at 0.6 kGy. Spoto et al. (1997)
irradiated orange juice concentrate and determined the effect of
storage time and temperature on juice quality and acceptability.
In that study, the highest dose ( up to 5.0 kGy ) combined with
25C storage resulted in loss of orange flavor and an
increase in bitter, medicinal , and cooked ratings by
sensory panelists. Lower dose (2.5 kGy) and cooler storage (0 or
5 C) were proposed as an acceptable processing regimen.
111
Introduction
Results
and Discussion
Table (15): Effects of irradiation on the sensory scores of pomegranate

Sensory
parameters
Storage
( days )
Irradiation dose
0
Color
Odor
Taste
Overall
acceptability
1.0
(kGy )
2.0
b
2.5
0
7
14
21
8.9 a0.05
7.2ab0.14
R
R
8.8 a0.05
8.2bb0.08
7.6bc0.08
R
8.2 a0.14
8.0cb0.03
7.2cc0.14
6.7cd0.08
8.2ba0.11
7.7db0.05
7.0dc0.05
6.4dd0.08
0
7
14
21
8.9aa0.03
3.8ab0.31R
R
R
8.8aa0.05
8.1bb0.16
3.8 bc0.15R
R
8.7ba0.08
8.3cb0.05
7.4cc0.14
3.6cd0.03R
8.6 ba0.05
8.3cb0.08
7.3cc0.14
3.5cd0.11R
0
7
14
21
8.8aa0.08
3.9ab0.31R
R
R
8.7aa0.05
7.2bb0.12
3.7bc0.11R
R
8.5ba0.06
7.5cb0.08
6.8cc0.05
3.7cd0.05R
8.4ba0.05
7.9db0.05
6.7cc0.12
3.5dd0.10R
0
7
14
21
8.8aa0.08
3.8ab0.11R
R
R
8.8aa0.11
7.6bb0.05
3.8bc0.08R
R
8.6ba0.08
7.9cb0.05
6.6cc0.15
3.8cd0.12R
8.6ba0.12
7.8cb0.08
6.3dc0.24
3.5dd0.11R
Where 0 (dislike extremely) to 9 (like extremely).

112
Introduction
Results
and Discussion
Odor
Color
TasteOverallacceptability
Fig. (14): Effects of irradiation on the sensory scores of pomegranate juice

during storage at 4C1. Where 0 (dislike extremely) to 9 (like
extremely).
113
Introduction
Results
and Discussion
Pickett et al. (2000) reported increasing off-flavor in

unpasteurized orange juice irradiated at 3.0 kGy, rending the
juice unpalatable. Niemira et al. (2001) found no evident
changes in the appearance or aroma of reconstituted orange
juice irradiated to 2.5 kGy. Song et al. (2007) found that the
overall sensory scores of the irradiated and non-irradiated
samples of vegetable juice were not significantly different
immediately after irradiation. However, the sensory quality of
the non-irradiated carrot and kale juice decreased with the
storage time, and it appeared to be the worst at storage of day 3.
Lee et al. (2009) observed no significant change in the sensory
scores in the irradiated (05 kGy) ready to use tamarind juices
when compared to that of non-irradiated juice they noticed a
strong off odor and off taste.
During storage, the sensory quality scores of nonirradiated and irradiated pomegranate juice samples decreased
but at different levels. The decline of sensory scores was higher
in non-irradiated pomegranate juice samples in comparison with
irradiated ones. After 7 days of storage the panelists rejected
non-irradiated (control) samples because of off-odor and color
changes. Thus, the control samples could not be used for
evaluation after 7 days of storage because of the deterioration in
quality and spoilage. Panelists rejected pomegranate juice
samples receiving 1.0, 2.0 and 2.5 kGy after 14, 21 and 21 days
of storage at 4C1, respectively.
114
Introduction
Results
and Discussion
4. Irradiation of fresh tomato juice

microbial load of tomato juice
Table (16) indicates that the non irradiated tomato juice
was highly contaminated with aerobic mesophillic bacteria,
lactic acid bacteria and yeasts and molds. The total log counts of
these microorganisms were 3.98, 3.67 and 3.57 cfu/ml,
respectively. The obtained microbial counts exceeded the level
of codex standard for tomato juice (Codex stan 49-1981). The
obtained high contamination levels of microorganisms was
expected and could be due to the high natural microflora of the
raw tomato, that come from soil, water or the hands of workers,
as well as the contamination during blending and packaging.
The obtained results were almost similar to those reported by
Prakash et al. (2002) who found that the diced tomato were
highly contaminated with mesophillic bacteria and mold and
yeasts at level of 4.40 and 4.58 Log cfu/ml, respectively. Hsu et
al. (2008) also found that the tomato juice were highly
contaminated with mesophillic bacteria, lactic acid bacteria and
mold and yeasts at the levels of 4.1, 4.2 and 3.7 log cfu/ml,
respectively. The high microbial counts found in tomato juice
would affect its quality and shelf-life.
Table (16) also shows the effect of different doses of
gamma irradiation (1.5, 3.0 and 4.5 kGy) on the total
mesophillic bacteria, lactic acid bacteria, mold and yeast
populations in tomato juice. Generally, irradiation caused a
significant decrease in all microbial counts under investigation
and the decrease was proportional with irradiation doses.
115
Introduction
Results
and Discussion

the microbial counts (log cfu/ml) contaminating tomato
juice
Microorganisms
Storage
( days )
0
Total
aerobic
mesophilic
bacteria
bacteria
0.0
1.5
3.0
4.5
3.98aa0.045
2.69ba0.002
1.93ca0.02
<10
5.86 b0.011
2.84 b0.007
b0.029
c
<10
5.27 c0.003
c0.01
dc0.006
15
6.86bd0.006
cd0.01
dd0.01
20
ce0.037
de0.001
25
5.79df0.34
3.67aa0.017
15
7.94 c0.02
10
10
5
Lactic acid
<10
<10
<10
<10
<10
<10
5.14 b0.045
6.96 c0.004
R
4.00 c0.025
b
6.54 d0.01
3.00 c0.01
<10
3.00 d0.006
3.96 d0.001
20
5.92 e0.014
4.14de0.017
25
6.47df0.004
3.57aa0.027
3.07ba0.02
ca0.008
da0.03
Total mold
5.63ab0.011
3.92bb0.007
cb0.01
db0.013
and yeasts
10
7.07ac0.03
6.00bc0.01
cc0.005
dc0.016
15
7.07bd0.03
cd0.005
dd0.008
20
ce0.01
6.71de0.019
25
df0.017

<10 = below detectable limit (<10 cfu/ml).
Mean values followed by different superscript (within rows) and different subscripts
(within columns) are significantly different (p < 0.05)
116
Introduction
Results
and Discussion
Irradiation dose of 1.5 kGy reduced the number of the

initial mesophillic bacteria by almost 1.3 log cycle. As well as,
it could be successfully reduce the lactic acid bacteria to below
the detectable counts (<10 cfu/ml). On the other hand,
irradiation at 1.5 kGy reduced total mold and yeasts by only 0.5
log cycles indicating that mold and yeasts are relatively more
resistance to gamma irradiation than bacteria. Fan et al. (2003b)
observed decay in Cilantro irradiated at 3.0 kGy during storage
and supported his observation that fungi are more resistant to
irradiation than bacteria. On the other hand, irradiation doses of
3.0 and 4.5 kGy were sufficient to reduce the number of the
initial molds and yeasts by 0.68 and 1.37, respectively.
Therefore, a dose of 3.0 and 4.5 kGy could be used to reduce
microbial counts of tomato juice to satisfactory level. Prakash
et al. (2002) reported that a dose of 3.07 kGy eliminated all
microorganisms contaminating diced tomatoes to no detectable
counts. Song et al. (2007) found that the irradiation doses of 1, 2
and 3 kGy could reduce the microbial populations of carrot juice
by 2.4, 3.8 and 5.9 log cycles, respectively and these irradiation
doses could reduce the microbial counts of kale juice by 1, 1.5
and 2.0 log cycles, respectively. These results indicate that the
microflora of the tomato, kale and carrot juice is fairly different,
so the radiation sensitivity of the microorganisms composing the
microflora should be considered in the radiation processing.
Table ( 16 ) indicated that during storage of tomato juice
at 4C 1, the viable counts of aerobic mesophillic bacteria,
lactic acid bacteria and mold and yeast in the non irradiated
tomato juice were progressively increased and reached 7.94,
6.96 and 7.07 log cfu/ml, respectively after 10 days. However,
these microorganisms reached almost similar counts in tomato
juice receiving 1.5 and 3.0 kGy but after 15 and 20 days,
respectively.
117
Introduction
Results
and Discussion
Total mesophillic aerobic bacterial count, lactic acid bacteria

and total mold and yeasts in tomato juice receiving 4.5kGy
reached 5.79, 6.47 and 7.34 log cfu/ml after 25 days of storage
at 4C 1. It is worthy to mention that at day 5 and 10 of
storage, these microbial counts in irradiated samples were
significantly less than of the unirradiated ones. Similar results
on microbial counts of irradiated (0.5, 1.24 and 3.70 kGy) diced
Roma tomatoes stored at 4C 1 have been shown by Song et
al. (2007).
4.2. Effect of irradiation and refrigeration storage on
pathogenic bacteria contaminating tomato juice
Table (17) shows the presence of coliform bacteria in
unirradiated tomato juice at level 2.4x102 MPN/ml. On the other
hand, Escherichia coli, Enterococcus faecalis and
Staphylococcus aureus were not detected in unirradiated tomato
juice. Coliform bacteria present in unirradiated tomato juice
were eliminated by the lowest irradiation dose used (i.e. 1.5
kGy) since the MPN was below detectable level (< 3) indicating
the sensitivity of coliforms to gamma radiation. During storage
of non irradiated tomato juice at 4C 1 there was progressive
and continuous increase in coliform bacteria, at day 5 and 10,
the MPN of coliform bacteria reached 2.1 x 104 and 4.3 x 104,
respectively. On the other hand, the MPN of coliform bacteria,
E. coli and other pathogens in the irradiated tomato juice
samples were below detectable level thought out the storage
period and were free from other pathogenic bacteria.
118
Introduction
Results
and Discussion

the some pathogens contaminating tomato juice.
Micro- organisms
Storage
( days )
0
5
10
15
20
25
0.0
2.4x102
2.1x104
4.3x104
R
R
R
1.5
<3
<3
<3
<3
R
R
3.0
<3
<3
<3
<3
<3
R
4.5
<3
<3
<3
<3
<3
<3
0
5
10
15
20
25
<3
<3
<3
R
R
R
<3
<3
<3
<3
R
R
<3
<3
<3
<3
<3
R
<3
<3
<3
<3
<3
<3
0
5
10
15
20
25
<100
<100
<100
R
R
R
<100
<100
<100
<100
R
R
<100
<100
<100
<100
<100
R
<100
<100
<100
<100
<100
<100
<100
0
<100
5
<100
10
Staphylococcus
R
15
aureus
R
( cfu / ml )
20
R
25
<100
<100
<100
<100
R
R
<100
<100
<100
<100
<100
R
<100
<100
<100
<100
<100
<100
Coliforms
( MPN/ml)
Escherichia coli
( MPN/ml )
Enterococcus
faecalis
( cfu / ml )
< 3 = No positive tubes have been shown in the first three dilutions.
<100 = below detectable level.
119
Introduction
Results
and Discussion

nutritional quality of tomato juice
The nutritional quality of irradiated fresh tomato juice in
comparison with unirradiated fresh tomato juice were
investigated by analyzing the contents of ascorbic acid (Vitamin
C) and lycopene pigment.
a) Ascorbic acids content
Fruit juices are an important source of vitamin C in the
total diet. However, ascorbic acid of fruit juices is readily
oxidized and lost during staying of the juices. Table (18) and
figure ( 20 ) shows that the ascorbic acid content of unirradiated
( control ) of tomato juice was 15.63 mg/100 ml. Similar result
have shown by Hsu et al. ( 2008 ) who found that ascorbic acid
in tomato juice was 18.60.8 mg/100 ml. The content of
ascorbic acid in tomato juice could vary according to the variety
of tomato, cultivar, plant origin, condition of planting,
harvesting, squeezing processetc. Generally, all irradiation
doses used significantly (p<0.05) decreased ascorbic acid
content of tomato juice and the decrease was proportional with
irradiation dose. This significant decrease of ascorbic acid is
apparent and could be due to the partial oxidation of ascorbic
acid to dehydroascorbic acid which not affecting tomato vitamin
C (vit. C = ascorbic acid + dehydroascorbic acid). This is
expected due to the fact that ascorbic acid is sensitive to
oxidation and gamma radiation cause water radiolysis of tomato
juice producing free radicals (H,OH,e ) and OH radicals are
strong oxidizing agent. Kilcast (1994) reported that ionizing
radiation can cause a partial conversion of ascorbic acid to
dehydroascorbic acid. So, it is useful in the next studies to
determine the amount of ascorbic acid and dehydroascorbic
acid. Graham and Stevenson (1997) found
120
Introduction
Results
and Discussion
that the dehydroascorbic acid content of tomato samples

increased immediately following irradiation. It should be
mentioned that both ascorbic acid and dehydroascorbic acid
have a vitamin C activity in the body.
During storage, ascorbic acid content of non-irradiated
and irradiated tomato juice were significantly decreased but at
different levels. After 10 days of storage at 4C1 ascorbic acid
content of non-irradiated samples decreased by 61.2%, while it
decreased in samples irradiated at 1.5, 3.0 and 4.5 kGy by 53.7,
24.9 and 25.5 %, respectively. It is clear that the percentage loss
of ascorbic acid content during storage of irradiated samples
was lower than that of non-irradiated samples and was dose
dependent. The loss of ascorbic acid in samples irradiated at 4.5
kGy was 38.6 % after 25 days of storage. It is worthy to mention
that both thermal treatments and high hydrostatic pressure
processing of fruit and vegetable juice as irradiation cause loss
in ascorbic acid but at different levels (Hsu et al., 2008 and
Song et al., 2007).
Table (18): Ascorbic acid content (mg/100 ml) of tomato juice treated
by irradiation during storage at 4C1
Storage
( days )

0
0
5
10
15
20
25
15.63aa0.03
6.50ab0.05
6.06ab0.03
R
R
R
1.5
13.23 a0.08
9.10bb0.05
6.13ac0.03
4.30bd0.05
R
R
b
3.0
6.53ca0.03
5.73cb0.03
4.90bc0.05
4.70bc0.05
4.60cc0.03
R
4.5
5.10 a0.05
4.46db0.03
3.80cc0.05
3.46cc0.02
3.16dc0.03
3.13dc0.01
d
R = Samples sensorially rejected, Mean values followed by different superscript

(within rows) and different subscripts (within columns) are significantly different (p <
0.05)
121
Introduction
Results
and Discussion
Fig. (15): Ascorbic acid content (mg/100 ml) of tomato juice treated by
irradiation during storage at 4C1.
b) Lycopene content
Most authors studied the thermal effect on lycopene
degradation but according to our knowledge no available data in
the literature dealing with the effect of ionizing radiation on the
lycopene content in foods. Therefore, one of the objectives of
this research was to investigate the effect of irradiation on
lycopene content of tomato juice. In this study, table (19) and
figure (21) show that the total lycopene content of nonirradiated (control) samples was 8.55 mg/100g wet weight. This
value are within the range (6.93-42.74 mg/100g wet weight)
reported for other tomato juices (Markovic et al., 2006). The
results indicated that irradiation at doses of 1.5, 3.0 and 4.5
kGy did not show significant effect on the total lycopene
content in tomato juice.
122
Introduction
Results
and Discussion
The results also show that the storage for 5 days resulted
in a significant decrease in the total lycopene content of both the
non irradiated and irradiated tomato juice samples, where the
loss reached 46.78, 20.42, 28.26 and 28.26% for samples
irradiated at 0.0, 1.5, 3.0 and 4.5 kGy, respectively. The data
revealed that the loss in total lycopene content of irradiated
samples were lower than that of non irradiated ones.
During further storage total lycopene content significantly
decreased in all samples (Table 19 and figure 21). Nguyen and
Schwartz (1999) mentioned in their review that processing and
storage of tomato products cause lycopene degradation. The
main causes of lycopene degradation are isomerization and
oxidation. It was suggested that the first stage of degradation is
the reversible isomeriztion of all trans-lycopene to less colored,
more oxidizable cis isomers (Boskovic, 1979).
Table ( 19 ): Lycopene content in irradiated and non- irradiated
tomato
juices (mg / 100 g wet weight ) during storage at
Storage
( days )
0
5
10
15
20
25
8.55aa0.005
4.55ab0.22
4.54ab0.20
R
R
R
Irradiation dose (kGy )

1.5
3.0
8.42aa0.02
6.70bb0.32
6.13bb0.18
5.12bc0.072
R
R
8.35aa0.21
5.99cb0.38
5.85cb0.005
5.83cb0.43
5.76cb0.43
R
4.5
8.35aa0.31
5.98cb0.45
5.83cb0.08
5.82cb0.09
5.14cc0.68
4.93dd0.45
4C1 R = Samples sensorially rejected.

123
Introduction
Results
and Discussion
Fig. (16): Lycopene content in irradiated and non- irradiated tomato

juices (mg / 100 g wet weight) during storage at 4C1.

physiochemical properties of tomato juice
a) Viscosity
The consistency of tomato juices expressed in viscosity
and level of syneresis (serum separation) is important for the
quality of tomato juices. The consistency of tomato products is
strongly affected by the composition of the pectins (Hsu et al.,
2008). Table (20) and figure (22) shows the effect of irradiation
on the viscosity of tomato juice, it is clear that the viscosity of
non irradiated (control) tomato juice was 462 cPs. irradiation
124
Introduction
Results
and Discussion
significantly decreased the viscosity of the tomato juice.

Irradiation at 1.5, 3.0 and 4.5 kGy significantly lowered the
viscosity from 46 to 38.0, 28.0 and 24.0 cPs, respectively.
The irradiation of pectin in aqueous solutions caused
degradation of macromolecules rather than the radiation induced
desertification of polysaccharide chains reveals the decrease in
viscosity of pectin solution (Zegota, 1999). The increase in
pectin methylesterase (PME) activity in response to irradiation
is in sharp contrast to the behavior for polygalacturonase (PG).
Increased PME activity was also reported for sweet cherries
irradiated at 2.0 and 5.0 kGy (Somogyi and Romani, 1964).
The same table and figure also show that the storage for 5
days at 4C1 resulted in a significant decrease in viscosity of
all tomato juice samples ( non irradiated and irradiated ), where
the viscosity was 17.0, 30.0, 24.0 and 22.0 cPs for samples
irradiated at 0.0, 1.5, 3.0 and 4.5 kGy, respectively. The
viscosity gradually decreased during further storage in all
samples.
Table (20): Viscosity (cPs) of tomato juices treated by irradiation
Storage
( days )
0
5
10
15
20
25

0
46.0aa2.0
17.0ab1.0
15.0ab1.0
R
R
R
1.5
38.0ba6.0
30.0bb2.0
25.0bc2.0
20.0bd2.0
R
R
3.0
28.0ca2.0
24.0cb2.0
22.0cc2.0
20.0bd2.0
18.0ce0.0
R
4.5
24.0da2.0
22.0db2.0
22.0cb2.0
20.0bc2.0
19.0cc1.0
14.0dd1.0
R = Samples sensorially rejected, Mean values followed by different superscript (within

rows) and different subscripts (within columns) are significantly different (p < 0.05).
125
Introduction
Results
and Discussion
Fig. (17): Viscosity (cPs) of tomato juices treated by irradiation during

storage at 4C1.
b) pH
Table (21) and fig (23) revealed that the pH of
unirradiated (control) tomato juice was 4.65 indicating acidic
medium. Irradiation at 1.5, 3.0 and 4.5 kGy showed no
significant effect on the acidity (pH) of the tomato juice. The
results also show that there were no significant differences in
pH as a result of irradiation and storage. Similarly, Prakash et
al. (2002) indicated that there were no significant differences in
the pH as a result of irradiation of diced tomato.
126
Introduction
Results
and Discussion
Table (21): pH value of irradiated and non-irradiated tomato juices

Storage
( days )

0
1.5
3.0
4.5
a
a
4.63 a0.01 4.63 a0.005
4.63aa0.01
0
4.64aa0.005 4.64aa0.01
4.64aa0.005
5
R
4.67aa0.005 4.68aa0.005
4.68aa0.005
10
R
4.73aa0.005 4.69aa0.005
4.69aa0.005
15
R
4.60aa0.005
4.62aa0.01
R
20
R
R
R
4.58aa0.005
25
4.65aa0.01
4.66aa0.01
Fig. (18): pH value of irradiated and non-irradiated tomato juice

during storage at 4C1.
127
Introduction
Results
and Discussion

sensory evaluation of tomato juice
Sensory evaluation of the tomato juice was performed in
the parameters of color, odor, taste, and overall acceptability.
Table (22) and figure (24) indicated that irradiation at doses of
1.5 and 3.0 kGy had no significant changes in the color, odor,
taste and overall acceptability. On the other hand, the highest
irradiation dose used (4.5 kGy) didnt affect the color of the
tomato juice significantly but significantly affect the odor, taste
and overall acceptability of tomato juice samples where the
panelists gave these samples the lowest scores. It is clear that all
irradiation doses used (1.5, 3.0 and 4.5 kGy) had no significant
effect. The result in table (19) confirm this fact as the irradiation
doses didnt effect lycopene content of tomato juice which is
responsible for the color of tomato.
Table (22) also indicated that during storage, the sensory
quality scores of non-irradiated and irradiated tomato juice
samples decreased but at different levels. The decline of sensory
scores was higher in non-irradiated tomato juice samples in
comparison with irradiated ones. After 10 days of storage the
panelists rejected non-irradiated (control) samples because of
off-odor, color changes and bad overall acceptability. Thus, the
panelists evaluated that the control samples could not be used
for further evaluation because of the deterioration in quality and
spoilage. Panelists rejected tomato juice samples receiving 1.5,
3.0 and 4.5 kGy after 15, 20 and 25 days of storage at 4C1,
respectively. Prakash et al. (2002) reported that sensory
evaluation of irradiated diced tomatoes indicated higher dosages
(1.24 and 3.70 kGy) of gamma irradiation impacted aroma,
flavor and textural properties and with increasing levels of
irradiation,
128
Introduction
Results
and Discussion
panelists noted a decrease in fresh tomato aroma and flavor and

an increase in ripe tomato aroma. Song et al. (2007) mentioned
that the overall sensory scores of the irradiated and nonirradiated samples of vegetable juice were not significantly
different immediately after irradiation. However, the sensory
quality of the non-irradiated carrot and kale juice decreased with
the storage time, and it appeared to be the worst at storage of
day 3.
129
Introduction
Results
and Discussion
Table (22): Effects of irradiation on the sensory scores of tomato juice

Sensory
parameters
Color
Odor
Taste
Overall
acceptability
Storage
( days )
Irradiation dose
( kGy )
0
5
10
15
20
25
0
8.8 a0.05
7.5ab0.05
4.0ac0.03
R
R
R
1.5
8.7 a0.05
8.3bb0.05
7.6bc0.15
3.9bd0.05R
R
R
3.0
8.7 a0.10
8.4bb0.05
7.6bc0.25
6.4cd0.05
3.8ce0.25R
R
4.5
8.6aa0.05
8.3bb0.05
7.5bc0.05
6.3cd0.05
4.0ce0.10
3.7df0.25R
0
5
10
15
20
25
8.7aa0.05
7.4ab0.05
3.6ac0.05R
R
R
R
8.6aa0.05
7.9bb0.10
6.7 bc0.25
4.0bd0.20R
R
R
8.6aa0.10
7.9bb0.05
7.1cc0.25
6.6cd0.15
4.0ce0.15R
R
8.0 ba0.05
7.2cb0.10
7.2cb0.45
7.1db0.55
6.6dc0.15
3.9dd0.1R
0
5
10
15
20
25
8.9aa0.05
7.7ab0.20
3.7ac0.25R
R
R
R
8.8aa0.05
7.9bb0.10
6.7bc0.20
3.6bd0.40R
R
R
8.7aa0.10
7.7cb0.20
7.2cc0.25
6.6cd0.10
3.9ce0.15R
R
8.2ba0.05
7.1db0.10
7.2cb0.15
7.1db0.15
6.3dc0.05
4.0dd0.05R
0
5
10
15
20
25
8.8aa0.05
7.7ab0.20
3.6ac0.30R
R
R
R
8.7aa0.05
7.9bb0.10
6.7bc0.20
3.9bd0.40R
R
R
8.7aa0.10
7.7cb0.20
7.2cc0.10
6.6cd0.10
3.9ce0.15R
R
8.1ba0.05
7.5db0.05
7.2cc0.05
7.2dc0.10
6.6dd0.15
3.8de0.10R
Where 0 (dislike extremely) to 9 (like extremely)

Mean values followed by different superscript ( within rows ) and different
subscripts ( within columns ) are significantly different ( p < 0.05 ).
130
Introduction
Results
and Discussion
Color
Odor
Taste
Overall acceptability
Fig. (19): Effects of irradiation on the sensory scores of tomato juice

during storage at 4C1. Where 0 (dislike extremely) to 9 (like
extremely).
131
Introduction
Results
and Discussion
5. Irradiation of fresh carrot juice

Microbiological evaluation of foods is of prime
importance due to the presence of spoilage microorganisms and
harmful human pathogens and their inactivation is essential to
ensure the hygienic quality of food material. Carrot juices
enjoys high consumer acceptance on account of its taste and
nutritional benefits. Carrot juice is however preferred as freshsqueezed. Although carrots are assumed to be safe, the
consumption of unpasteurized juices may be dangerous since
they could be a potential source of bacterial pathogens (Ryu
and Beuchat, 1998). In our study table (23) shows that the
original microbial population (aerobic mesophillic bacteria,
lactic acid bacteria and yeasts and molds) of carrot juices were
4.98, 4.76 and 3.25 log cfu/ml, respectively. Song et al. (2007)
found that the initial population of total aerobic bacteria in the
carrot juice were 5.96 log cfu/ml. This levels is higher than that
obtained in our study. This could be due to the differences in
varieties, environment in which plants are grown, post-harvest
handling, processing conditions, etc.
The results of the viable counts of aerobic mesophillic
bacteria, lactic acid bacteria and yeasts and molds in carrot juice
after irradiation treatment (1.5, 3.0 and 4.0 kGy) are presented
in table (23). The results in table (23) also indicate that gamma
irradiation significantly reduced the microbial contamination
level with a dose dependent tendency (p<0.05).
132
Introduction
Results
and Discussion

the microbial counts (log cfu/ml) contaminating carrot
juice
Microorganisms
Storage
( days )

0.0
1.5
3.0
4.0
Total
aerobic
mesophilic
bacteria
0
3
5
10
4.98aa0.01
7.10ab0.03
R
R
3.92ba0.01
4.74bb0.02
6.20bc0.06
7.85bd0.005
2.91ca0.13
3.69cb0.005
4.68cc0.01
7.03cd0.03
<10
2.69db0.005
3.73dc0.04
6.05dd0.05
Lactic acid
bacteria
0
3
5
10
4.76aa0.005
6.89ab0.005
R
R
2.90ba0.09
4.98bb0.01
5.03bc0.03
7.78bd0.01
2.48ca0.12
2.48cb0.015
4.02cc0.02
7.34cd0.04
<10
<10
2.75dc0.15
4.02dd0.02
0
3
5
10
3.25aa0.05
5.17ab0.03
R
R
1.86ba0.09
2.25bb0.025
3.66bc0.02
4.52bd0.16
1.73ca0.04
1.53cb0.06
2.85cc0.01
3.40cd0.01
<10
<10
<10
<10
Total
molds and
yeasts

133
Introduction
Results
and Discussion
The levels of 1.06, 1.86 and 1.39 log cycle reduction of

total aerobic bacterial counts, lactic acid bacteria and mold and
yeast were achieved by 1.5 kGy, respectively. 3.0 kGy could
reduce them by 2.07, 2.28 and 1.52, respectively. On the other
hand, a dose of 4.0 kGy could be successfully reduce them to
below the detectable counts (<10 cfu/ml). Chaudry et al. ( 2004
) reported that the irradiation doses of 0.5, 1.0 reduce the the
number of the initial total bacterial count of carrots by 0.83 and
0.29 log cycles, respectively and doses 2.0, 2.5 and 3.0 kGy
eliminate the total bacterial count. The same doses 0.5, 1.0, 2.0,
2.5 and 3.0 kGy eliminate the total fungal counts completely of
carrots. Song et al. (2007) found that the irradiation doses of 1
and 2 kGy could reduce the microbial populations of carrot juice
by 2.4 and 3.8 log cycles, respectively. They added that all the
aerobic bacteria and coliform in the fresh carrot juice were
eliminated by irradiation at 3 kGy. However, radiation doses up
to 5 kGy could not completely eliminate the bacteria in the fresh
kale juice indicating that the microflora of the carrot and kale
juice is fairly different. Chervin and Boisseau (1994) reported
that the growth of aerobic and lactic microflora on shredded
carrots was inhibited by irradiation at 2.0 kGy and chlorination
treatment, and the sensory analysis panellists preferred the
irradiated vegetables.
The effect of refrigeration storage on the microbial
populations (aerobic bacteria, lactic acid bacteria, molds and
yeasts) of non- irradiated and irradiated carrot juice samples is
also tabulated in table (23). It is evident from the results in the
table that all microbial populations are significantly increased as
storage period increased but the rate of increase was lower in
irradiated samples in comparison with non-irradiated ones. The
counts of total aerobic bacteria, lactic acid bacteria, molds and
134
Introduction
Results
and Discussion
yeasts in non-irradiated carrot juice were increased and reached

7.1, 6.89 and 5.17 log cycles after 3 days, respectively.
The counts of these microbial populations in irradiated
carrot juice samples at 1.5 and 3.0 kGy were also increased and
reached almost the same counts but after 10 days of storage.
Total aerobic bacterial counts and lactic acid bacteria of carrot
juice samples receiving 4.0 kGy gradually increased during
storage at 4C1 reaching 6.05 and 4.0 log cycles after 10 days.
However, total yeasts and molds were below detectable level
throughout the storage period. These results show that 3 or 4
kGy irradiation doses could significantly reduce the microbial
populations and extended the shelf life of the fresh carrot juice
to 10 days at 4C1 from the view point of microbial counts.
Farkas et al. (1997) showed that 1 kGy dose of gamma
radiation was sufficient to reduce bacterial load, improve
microbiological shelf life and extend sensorial quality of carrots.
Microbial spoilage of non-irradiated carrot juice proceeds
rapidly because the initial total viable count is generally high
(typically 105-106 cfu/ml) (Park et al., 2002). Song et al. (2007)
reported that total aerobic bacteria count reached 9.09 log
cfu/ml after 3 days of storage in carrot juice.
Results (table 24) revealed the presence of coliform
bacteria (460 MPN/ml), Escherichia coli (21 MPN/ml) and
Enterococcus faecalis (3.5x102 cfu/ml) indicating that a serious
health hazard due to the possible presence of enteric pathogens.
Like non pathogenic microorganisms, pathogens come from
the environment in which plant are grown, as well as from post
135
Introduction
Results
and Discussion
harvest handling, wash water, processing, etc. The entry of such

pathogens subsequently in fresh squeezed juice could be
facilitated if damaged or partially carrots are used (Beuchat,
1996). Ghosh et al. (2004) found high numbers of fecal and
total colifroms ( 104 and 107 cfu/g ) in carrot destined for
juicing. In contrast, Staphylococcus aureus and Aeromonas
hydrophila were not detected in our unirradiated carrot juice.
This implies that unprocessed vegetable juice is a harbor for
human pathogenic bacteria, resulting in a foodborne outbreak.
Therefore, vegetable juice should undergo some type of
processing to inactivate most of the microorganisms. The same
table also shows the inactivation effects of -irradiation on
pathogens found in carrot juice. A dose of 1.0 kGy or above
effectively reduced the coliform and Escherichia coli
populations to below the detectable levels (< 3), but was not
enough for elimination of
Enterococcus faecalis. While,
irradiation dose of 3 or 4 kGy was effectively for elimination of
Enterococcus faecalis found in carrot juice. Also, Alighourchi
et al. (2008) showed that low-dose gamma radiation (0.5 kGy)
treatment potentially could be eliminating Escherichia coli
without any detectable effect on the organoleptic quality of the
product.
During storage of carrot juice at 4C1, total coliforms
and E.coli decreased from 460 and 21 MPN/ml to 150 and < 3
MPN/ml, respectively at day 3. Meanwhile, Enterococcus
faecalis increased reaching 7.6x103 cfu/ml. MPN of coliform
bacteria, E.coli and Enterococcus faecalis in all irradiated
carrot juice samples were below detectable level thoughout the
refrigeration storage period.
136
Introduction
Results
and Discussion
Table (24): Effect of -irradiation and subsequent storage ( 4C1 ) on

the some pathogens contaminating carrot juice
Microorganisms
Coliforms
( MPN/ml)
Escherichia coli
( MPN/ml )
Enterococcus
faecalis
( cfu / ml )
Staphylococcus
aureus
( cfu / ml )
Aeromonas
hydrophila
( cfu / ml )
Storage
( days )

0.0
1.5
3.0
4.0
0
3
5
10
460
150
R
R
<3
<3
<3
<3
<3
<3
<3
<3
<3
<3
<3
<3
0
3
5
10
21
<3
R
R
<3
<3
<3
<3
<3
<3
<3
<3
<3
<3
<3
<3
0
3
5
10
3.5x102
7.6x103
<100
R
R
2.0x102
<100
<100
<100
<100
<100
<100
<100
<100
<100
<100
<100
0
3
5
10
<100
<100
R
R
<100
<100
<100
<100
<100
<100
<100
<100
<100
<100
<100
<100
0
3
5
10
<100
<100
R
R
<100
<100
<100
<100
<100
<100
<100
<100
<100
<100
<100
<100

<100 = below detectable level
137
Introduction
Results
and Discussion
5.3. Effect of -irradiation and refrigeration storage on the

nutritional quality of carrot juice
a) Ascorbic acid content
Ascorbic acid content (AA) was quantified in all the
products, since it is the natural form presented in fruits and
vegetables.Table (25) shows that unirradiated carrot juice
samples contained 8.8 mg/100g of AA. Song et al. (2007) found
that ascorbic acid content in carrot juice was 4.37 mg/100 ml
indicating variation in ascorbic acid content of carrot juices. The
variety of carrot, cultivar, plant origin and condition of planting.
All of these factors affect the variation on the content of
ascorbic acid in carrot juice.
irradiation at dose of 1.5 kGy did not show a significant
changes in the AA content, while doses of 3.0 and 4.0 kGy
resulted in 25.79% and 55.11% loss of total AA, respectively
indicating that these doses significantly ( p<0.05) decreased
ascorbic acid content of carrot juice. A.A is the most sensitive
water-soluble vitamin to irradiation (Kilcast, 1994) and is
highly sensitive to various modes of degradation (Tannenbaum
et al., 1985). This significant decrease of ascorbic acid is
apparent and could be due to the partial oxidation of ascorbic
acid to dehydroascorbic acid which not affecting carrot juice on
vitamin C. Song et al. (2007) reported that irradiation resulted
in a dose dependent reduction of the ascorbic acid content in
carrot and kale juice, however, the contents of the total ascorbic
acid, including dehydroascorbic acid, were stable up to 3 kGy of
irradiation. Also, Wen et al. (2006) observed that the vitamin C
content was decreased following the increasing doses of gamma
irradiation in lyceum fruit. The reduction of the vitamin C
content
138
Introduction
Results
and Discussion
should not be a significant problem, because carrot juice is the

essential source of B-carotene not Vitamin C.
During refrigeration storage, there was a decrease in the
ascorbic acid content of carrot juice but at different levels
(Table 25). After 3 days of storage at 4C 1, ascorbic acid
content decreased by 4.54% in 0.0 kGy, while it decreased in
samples irradiated at 1.5, 3.0 and 4.0 kGy by 1.53 %, 20.82%
and 21.01%, respectively. It is obvious that 1.5 kGy showed the
least loss in ascorbic acid content during storage. Almost similar
results was obtained by Song et al. (2007) who found that 3.0
kGy caused a loss by 35.12% in ascorbic acid content of carrot
juice after 3 days storage.
Table (25): Ascorbic acid content (mg/100 ml) of carrot juice treated
with irradiation during storage at 4C1
Storage
( days )
0
3
5
10

0
8.80aa1.30
8.40aa0.03
R
R
1.5
8.45aa0.65
8.32aa0.05
8.20ba1.05
8.05ba0.20
3.0
6.53ba0.05
5.17bb0.03
3.85cc0.05
3.82cc0.02
4.0
3.95ca0.05
3.12cb0.03
2.65dc0.05
2.46dc0.04

subscripts ( within columns ) are significantly different ( p < 0.05 )
139
Introduction
Results
and Discussion
Fig . (20): Ascorbic acid content (mg/100 ml) of carrot juice treated
with irradiation during storage at 4C1.
b) Total carotenoids content

Carotenoids are among the most important nutrients in
food, owing to their diverse functions and actions. The attractive
red or yellow colour conferred to many foods is their most
apparent contribution to food quality. It is accepted that
carotenoids in general and carotenes in particular provide
significant antioxidant activity to the human food and animal
feed supply, and thus may be responsible for some of the
significant correlations between increased intake of vegetables
providing significant carotenoid content and improved health
status (Akhtar and Bryan, 2008). Recently it has been reported
that the world carotenoid market is expected to reach U.S. $1.06
billion by the end of 2010 as consumers continue to look for
natural ingredients (Nutraingredient, 2007).
140
Introduction
Results
and Discussion
Total carotenoids in our irradiated and non irradiated

carrot juice are presented in table (26). It is evident that the total
carotenoids in non irradiated carrot juice were 3738 g/g. This
results was in the range of total carotenoids content in carrot
samples prepared in large quantities (expressed on insoluble
solids basis), as analysed by spectrophotometry (2237- 4714
g/g) reported by (Sant'Ana et al., 1998). The data revealed
that no significant difference in total carotene content between
irradiated samples and non irradiated. Gamma-irradiation did
not seem to decrease the total carotenoids content. According to
the International Consultative Group on Food Irradiation
(ICGFI), gamma-irradiation does not affect the content of bcarotene or other carotenoids with pro-vitaminic A activity
(ICGFI, 1999). Also, Wen et al. (2006) reported that gamma
irradiation had no effect on b- carotene content of Lycium fruit.
During the refrigeration storage period, the total
carotenoids decreased significantly in all samples. It is well
known that carotenoids are extremely susceptible to
degradation. Their highly unsaturated structure makes them
sensitive to heat, oxygen and light (Britton, 1992).
141
Introduction
Results
and Discussion
Table (26): Effect of irradiation and refrigeration storage on the

total carotenoids (g/g) of carrot juice
Storage
( days )
0
3738aa38.5
1.5
3535aa20.0
3.0
3451aa10.0
4.0
3400aa20.0
2075ab28.0
3535ba35.0
2927cb23.2
1282db87.50
10
1360bb40.0
1325bc75.0
1250bb50.0

Fig. ( 21 ): Effect of irradiation and refrigeration storage on the

total
carotenoids of carrot juice.
142
Introduction
Results
and Discussion

a) Viscosity
The rheological behaviour of juices is largely influenced
by their quantitative and qualitative composition and, therefore,
it will depend on the fruit type and on the treatments to which it
is subjected during processing. Table (27) indicated the
viscosity of carrot juice, it is clear that all irradiation doses used
(1.5, 3.0 and 4.0 kGy) had no significant decrease in the
viscosity of carrot juice. However, a significant increase in the
viscosity of the juice has observed during refrigeration storage,
and this may be due to the increase in the microbial load of the
juice which leads to deterioration of the juice and increase its
viscosity. Also Huis int Veld (1996) reported that chemical
spoilage processes of food may also bring about physical
changes such as increased the viscosity, gelation, sedimentation
or color change of food.
Table (27): Viscosity (cPs) of carrot juices treated by irradiation
Storage
( days )
0
4.0aa0.005
4.0aa0.02
1.5
4.0aa1.0
3.0
4.0
4.0aa2.0
12.0ab0.005
4.0ba0.01
4.0ba1.0
4.0ba2.0
9.0bb1.0
7.0cb0.5
6.0db0.01
10
18.0bc2.0
17.0cc3.0
16.0dc0.02

143
Introduction
Results
and Discussion
Fig. (22): Viscosity (cPs) of carrot juices treated by irradiation during

storage at 4C1.
b) pH
The pH of carrot juice ( control ) was 6.92 indicating a
neutral medium, irradiation at 1.5, 3.0 and 4.0 kGy showed no
significant effect on the pH of the carrot juice ( table 28 ).
However, during storage the pH of non-irradiated and irradiated
carrot juice samples significantly decreased. Wang et al. (2006)
studying the kinetics of enzymatic darkening in carrot juice
during storage, observed a considerable reduction in the pH over
time for samples stored at 25 and 37 C, which was not noted at
-18 and 0 C. The authors assigned the decrease in the total
sugars, total amino acids and pH to the Maillard reaction. They
attributed the pH reduction to the formation of
hydroxymethylfurfural (HMF) from the reactions involving
amino acids and reducing sugars. The darkening of the juice was
also attributed to HMF formation. The physico-chemical results
for untreated juice are in
144
Introduction
Results
and Discussion
agreement with Saldana et al. (1976), Lombrana and Dias

(1985), Bawa and Saini (1987), Anastasakis et al. (1987).
Table (28): pH value of irradiated and non-irradiated carrot juices
Storage
( days )
0
0
3
5
10
6.92aa0.04
5.37ab0.21
R
R
1.5
6.85aa0.005
6.85ba0.005
5.52bb0.125
4.62bc0.20
3.0
6.53aa0.01
6.52ba0.01
6.45ca0.065
5.51cc0.005
4.0
6.77aa0.07
6.78ba0.07
6.98ca0.13
5.52cc0.005

subscripts ( within columns ) are significantly different ( p < 0.05 ).
Fig. ( 23 ): pH value of irradiated and non-irradiated carrot juices

145
Introduction
Results
and Discussion
5.5. Effect of -irradition and refrigeration storage on the

All the carrot juice samples (irradiated and nonirradiated)
were subjected to sensory evaluation for color, odor, taste and
overall acceptability using 9-hedonic scale. The panellists scores
were tabulated in table (29) and illustrated in figure (29).
Irradiation at 1.5 and 3.0 kGy had no significant (p<0.05) effect
on sensory characteristics of carrot juice as the panellists gave
these samples almost the same scores. However, irradiation dose
of 4.0 kGy significantly (p<0.05) affected the sensory
characteristics of carrot juice. This suggests that carrot juice
should not be irradiated at more than 3.0 kGy doses for storage
at refrigeration temperature as 4C1.
During refrigeration storage at 4C1, the sensory
characteristics of all carrot juice samples (irradiated or not)
significantly decreased. Odor, taste and overall acceptability
scores became the worst at day 3, where the panellists rejected
these samples because of off-odor and very bad taste. Therefore,
control samples could not be used for evaluation after that time.
It is clear that carrot juice had very short shelf-life, not more
than 2 days at refrigeration temperature. Similar results have
been found by Song et al. (2007) who reported that the sensory
quality of non irradiated carrot juice (control) was deteriorated
at day 3. The panellists rejected carrot juice samples receiving
1.5 kGy after 5 days of refrigeration storage. Meanwhile, carrot
juice samples irradiated at 3 kGy were sensorial rejected after
10 days or refrigerated storage. Carrot juice samples exposed to
4 kGy still accepted at day 9. This means that irradiation doses
at 1.5, 3.0 and 4.0 kGy could extend the shelf-life of carrot
samples at refrigeration temperature to 4, 8 and 9 days,
respectively. It could
146
Introduction
Results
and Discussion
be concluded that irradiation dose of 3.0 kGy could be used to

extend the shelf life of carrot juice without adverse effect on
sensory quality.
Table (29): Effects of irradiation on the sensory scores of carrot juice
Sensory
parameters
Storage
( days )
Irradiation dose
0
color
odor
Taste
Overall
acceptability
( kGy )
1.5
3.0
a
4.0
b
0
3
5
10
8.9 a0.05
8.6ab0.05
R
R
8.8 a0.05
8.6ab0.05
7.5bc0.15
6.7bd0.05
8.8 a0.10
8.6ab0.05
7.5bc0.25
6.8bd0.05
8.5 a0.05
8.4ba0.05
7.4bb0.05
6.8bc0.05
0
3
5
10
8.7aa0.05
3.8ab0.05R
R
R
8.6aa0.05
7.3bb0.10
3.9 bc0.25R
R
8.6aa0.10
7.7cb0.05
6.7cc0.25
3.8cd0.15R
8.4 ba0.05
7.7cb0.10
6.3dc0.45
3.9cd0.55R
0
3
5
10
8.9aa0.05
3.7ab0.20R
R
R
8.8aa0.05
6.5bb0.10
3.3bc0.20R
R
8.8aa0.10
7.5cb0.20
6.3cc0.25
3.6cd0.10R
8.6ba0.05
7.7db0.10
6.5cc0.15
3.3cd0.15R
0
3
5
10
8.7aa0.05
3.7ab0.20R
R
R
8.6aa0.05
6.5bb0.10
3.5bc0.20R
R
8.6aa0.10
7.5cb0.20
6.7cc0.10
3.6cd0.10R
8.5ba0.05
7.8db0.05
6.8cc0.05
3.9cd0.10R

subscripts (within columns) are significantly different ( p < 0.05 )
147
Introduction
Results
and Discussion
Color
Odor
Taste
Overall acceptability
Fig. (24): Effects of irradiation on the sensory scores of carrot juice

148
Introduction
Results
and Discussion
6.
Combination
cinnamaldehdye
treatment
of
irradiation
and
6.1. Effect of irradiation and cinnamaldehdye on the

Radiation treatment is a process with excellent potential
for controlling or eliminating food borne pathogens in food.
However, the use of radiation to kill pathogens is limited
because radiation induces effects in the sensory quality of the
food products (Lacroix et al., 1991). When irradiation is used in
combination with other preservation methods such as addition
of spices and plant extracts, the global efficiency is reinforced
through synergistic action, and the irradiation doses can be
reduced without affecting the product quality (Mahrour et al.,
1998). Essential oils are known as antimicrobial agents that
could be used to control food spoilage and food borne
pathogenic bacteria (Espina et al., 2011). Preservation and shelf
life extension have been the historical focus of research on
irradiation of juices (Niemira and Deschenes, 2004).
The results of previous study indicate that although
irradiation dose of 4.0 kGy reduced all microbial counts to
below detectable levels thereby improve microbial shelf life and
safety, it had an adverse effect on sensory quality of carrot juice.
While, irradiation dose of 3.0 kGy could reduce the microbial
counts to great extent as well as it reduced coliform bacteria,
Escherichia coli and Enterococcus faecalis to below detectable
levels without adverse effects on sensory qualities of carrot
juice. But carrot juice samples irradiated at this irradiation dose
had limited shelf-life at refrigeration temperature (4C1)
almost 8 days. To get more refrigeration shelf-life extension of
carrot juice, it was designed to use irradiation in combination
with other preservation techniques.
149
Introduction
Results
and Discussion
Table (30) show the effect of -irradiation (3 kGy ) in

combination with cinnamaldehyde ( 8l/100ml) on total aerobic
bacterial counts, lactic acid bacteria and total mold and yeasts
of carrot juice samples during refrigeration storage. Addition of
cinnamaldehyde alone at concentration of 8l/100ml reduced
the counts of these microorganisms by 1.01, 0.23 And 0.5 log
cycles,
respectively.
Meanwhile,
combination
of
cinnamaldehyde at 8l/100ml concentration and irradiation at
3.0 kGy reduced all microbial counts under investigation to
below detectable levels <10 cfu/ml. Conner and Beuchat
(1984) proposed that essential oils damage a variety of enzyme
systems of yeasts affecting structural component synthesis and
energy production. They also reported that essential oils prohibit
repair of injury in yeasts metabolically or structurally damaged
as a result of being exposed to elevated temperatures. Oussalah
et al. (2006) reported that the mechanism of action of the
cinnamon oil and its main active compound on microorganisms
appear to be related to the cell membrane from a slight
disruption seem to occur, causing dispersion of the proton
motive force by leakage of small ions without leakage of large
cell components, such as ATP, which were subsequently
degraded by the ATPase enzyme. This fact could be attributed
to the hydrophobic nature of the oil, which may directly diffuse
across the lipid bilayer of the cell membrane, and /or gain acess
to the periplasm and deeper parts of the cell through the porins,
which have also showed to allow the penetration of lipophilic
substances of Low molecular weight at significant rates, despite
of its hydrophilic nature (Helander et al., 1998).
150
Introduction
Results
and Discussion
Table (30): Effect of -irradiation and cinnamaldehyde (8l/100ml) on

the microbial counts (log cfu/ml) contaminating carrot
Microorganisms
Total
aerobic
mesophilic
bacteria
Lactic acid
bacteria
Total mold
and yeasts
Storage
( days )
Treatments
0.0
8l
cinnamaldehyde
3.0 kGy + 8l
cinnamaldehyde
0
3
7
14
21
28
4.25aa0.01
7.11ab0.03
R
R
R
R
3.24ba0.01
4.33bb0.02
6.40bc0.01
R
R
R
<10
2.48cb0.12
3.32cc0.05
4.69cd0.05
6.41ce0.02
7.53cf0.02
0
3
7
14
21
28
2.20aa0.03
4.98ab0.01
R
R
R
R
1.97ba0.05
3.98bb0.01
4.02bc0.02
R
R
R
<10
<10
<10
<10
<10
<10
0
3
7
14
21
28
3.15aa0.05
5.15ab0.03
R
R
R
R
2.65ba0.01
3.25bb0.02
5.40bc0.01
R
R
R
<10
2.18cb0.12
3.03cc0.03
4.96cd0.08
5.73ce0.04
6.94cf0.04

151
Introduction
Results
and Discussion
During storage, it is evident from the results in the table

(30) that all microbial populations are significantly increased as
storage period increase but the rate of increase was lower in the
samples contain 8l/100ml cinnamaldehdye and was more
lower in the samples contain 8l/100ml cinnamaldehdye and
irradiated at 3.0 kGy. The counts of total aerobic bacteria, lactic
acid bacteria and yeasts in non-irradiated carrot juice were
increased and reached 7.11, 4.98 and 5.15 log cycles after 3
days of storage at 4C1, respectively. The counts of these
microbial populations in carrot juice containing 8l/100ml
cinnamaldehdye and carrot juices containing 8l/100ml
cinnamaldehdye and irradiated at 3.0 kGy were also increased.
These counts almost reached almost the same counts but after 7
days of storage for samples containing 8l/100ml
cinnamaldehdye, and 28 days for samples containing 8l/100ml
cinnamaldehdye in addition to irradiation at 3.0 kGy. This
indicated that combination treatment of irradiation (3.0 kGy)
and 8l/100ml cinnamaldehdye increased the shelf life of carrot
juice from 3 days to 28 days from the view point of microbial
counts. Valero and Salmeron (2003) reported that the addition
of 5l cinnamon essential oil to 100 ml of carrot broth,
inoculated with Bacillus cereus spores and preserved at
temperature of 8C or lower, made it possible to inhibit its
germination and growth for at least 60 days in a model RMP
(refrigerated minimally processed foods) food made with carrots
at 16C.
Results (Table 31) revealed the presence of coliform
bacteria (350 MPN/ml), Escherichia coli (22 MPN/ml). The
table shows that no reduction effects of 8l/100ml
cinnamaldehdye on total coliform bacteria of carrot juice, while
it was slight
152
Introduction
Results
and Discussion
reduction in E.coli. Meanwhile, combination treatment of

irradiation (3.0 kGy) and 8l/100ml cinnamaldehyde was
effectively for elimination of coliform bacteria and E.coli
populations found in carrot juice below the detectable levels (<
3).
Lacroix et al. (2009) showed that irradiation technology
in combination with other treatments is effective for reducing or
eliminating the growth of bacteria and subsequently extending
the shelf life of readytoeat vegetables, by increasing the
radiosensitization of bacteria and by lowering the dose
necessary to eliminate them. Abd El-Aziz and Abd El-Khalek
(2010) reported that the cinnamon essential oils showed
moderate effects against Zygosaccharomyces rouxii,
Enterococcus faecalis and Escherichia coli. Also they added
that combination treatment of 4 or 6 l/ml of orange peel
essential oil in orange juice and irradiation at 2.0 or 3.0 kGy
completely inhibited the growth of these microorganisms after
96 h.
153
Introduction
Results
and Discussion
Table (31): Effect of -irradiation and cinnamaldehyde ( 8l/100ml )

on pathogens contaminating carrot juice during storage at 4C
Micro- Storage
organisms ( days )
Treatments
0.0
8l
Coliforms
(MPN/ml)
0
7
14
21
28
350
112
R
R
R
350
112
R
R
R
3.0 kGy + 8l
cinnamaldehyde
<3
<3
<3
<3
<3
Escherichia coli
( MPN/ml )
0
7
14
21
28
22
<3
R
R
R
19
<3
R
R
R
<3
<3
<3
<3
<3
0
7
14
21
28
<100
<100
R
R
R
<100
<100
R
R
R
<100
<100
<100
<100
<100
0
7
14
21
28
<100
<100
R
R
R
<100
<100
R
R
R
<100
<100
<100
<100
<100
0
7
14
21
28
<100
<100
R
R
R
<100
<100
R
R
R
<100
<100
<100
<100
<100
cinnamaldehyde
Enterococcus
faecalis
( cfu/ml )
Staphylococcus
aureus
( cfu/ml )
Aeromonas
hydrohila
(cfu/ml)
;
<100 = below detectable level.
154
Introduction
Results
and Discussion

a) Viscosity
Table (32) indicated the viscosity of carrot juice, it is
clear that all treatments had no significant decrease in the
viscosity of carrot juice. However, a significant increase in the
viscosity of the juice has observed during refrigeration storage,
and this may be due to the increase in the microbial load of the
juice which leads to deterioration of the juice and increase its
viscosity. While no increase occurs in the irradiated juice
containing 8l/100ml cinnamaldehdye.
Table (32): Viscosity (cPs) of carrot juices treated by irradiation in
combination with cinnamaldehyde (8l/100ml)
Storage
( days )
8l cinnamaldehyde
3.0 kGy + 8l
cinnamaldehyde
4.0aa0.001
4.0aa0.01
4.0aa0.003
12.0ab0.002
10.0bb0.05
4.0ca0.002
14
4.0ca0.005
21
28
R
R
R
R
4.0ca0.001
4.0ca0.002

155
Introduction
Results
and Discussion
Fig. (25): Viscosity (cPs) of carrot juices in combination with

cinnamaldehyde (8l/100ml).
b) pH
The pH of carrot juice (control) was 6.98 indicating a
neutral medium. (Table 33 and fig 31) showed no significant
effect on the pH of the carrot juice as a result of adding
cinnamaldehyde alone or in combination with irradiation.
However, during storage the pH of control sample and the
sample containing 8l/100 ml cinnamaldehyde significantly
decreased. While, the irradiated sample with 8l/100 ml
cinnamaldehyde showed no significant decrease in the pH of the
juice during storage. Baskaran et al. (2010) reported that
addition of Trans-cinnamaldehyde didnt result in any
significant change in the apple juice or apple cider pH.
156
Introduction
Results
and Discussion
Table (33): pH values of carrot juices treated by irradiation in

combination with cinnamaldehyde (8l/100ml)
Storage
( days )
8l cinnamaldehyde
3.0 kGy + 8l
cinnamaldehyde
6.98aa0.04
7.0aa0.01
6.99aa0.01
3.05ab0.01
3.89ab0.02
6.30ba0.01
14
6.29ba0.01
21
28
R
R
R
R
6.07ba0.01
6.04ba0.01

Fig. (26): pH values of carrot juices treated by irradiation in

combination with cinnamaldehyde ( 8l/100ml).
157
Introduction
Results
and Discussion
6.4. Effect of irradiation and cinnamaldehdye on sensory

evaluation of carrot juice
The evaluations of the panellists are summarized in table
(34). There was no significant effect on the color, odor, taste
and overall acceptability of juices as a result of adding
cinnamaldehyde alone or in combination with irradiation.
Although the essential oil of cinnamon has a strong smell and
flavour,the
organoleptic
and,especially,
smell-taste
characteristics of carrot juice seem to mask any perceived
differences among the samples. Similar results was indicated by
Valero and Salmeron (2003) who found that the sensory
analysis related to the use of cinnamon essential oil as an
antimicrobial agent in carrot broth showed a masking effect and
provoked smell-taste reactions in the panel members such as
not bad or I liked it a lot.
During storage at 4C1, the sensory scores significantly
decreased in all the samples but at different rate. The rate of
decrease in carrot juices containing 8l/100 ml cinnamaldehyde
and irradiated at 3.0 kGy was the lowest in comparison with
other treatments. The panellists rejected carrot juice samples
(control) at day 3 due to the off-odor and bad taste. Carrot juice
samples containing cinnamaldehyde were organoleptically
rejected after 7 day of storage at 4C1. Combination treatment
of cinnamaldehyde and irradiation at 3.0 kGy extended the
shelf-life of carrot juice to 21 days from the view point of
organoleptic quality.
158
Introduction
Results
and Discussion
Table (34) Effect of -irradiation and cinnamaldehyde ( 8l/100ml ) on

sensory evaluation of carrot juice during storage at 4C
Sensory
parameters
Storage
( days )
Treatments
0
Color
Odor
Taste
Overall
acceptability
8l cinnamaldehyde
.
0
3
7
14
21
28
8.7aa0.05
8.6aa0.05
R
R
R
R
8.6aa0.05
8.5aa0.05
8.4 bb0.05
R
R
R
8.6aa0.10
8.6aa0.05
8.5ba0.25
8.5ca0.05
8.4cb0.25
8.3cb0.25
0
3
7
14
21
28
8.7aa0.05
4.0ab0.05R
R
R
R
R
8.7aa0.05
6.8bb0.10
3.7 bc0.1R
R
R
R
8.5ba0.10
8.3cb0.05
7.7cc0.25
6.6cd0.15
6.3ce0.15
3.9 cf0.15R
0
3
7
14
21
28
8.7aa0.05
3.8ab0.20R
R
R
R
R
8.5 ba0.05
6.3bb0.10
3.8bc0.20R
R
R
R
8.4ba0.10
8.1cb0.20
7.6cc0.25
6.8cd0.10
6.4ce0.15
3.8 cf0.15R
0
3
7
14
21
28
8.7aa0.05
3.9ab0.20R
R
R
R
R
8.5aa0.05
6.15bb0.10
3.9bc0.20R
R
R
R
8.4aa0.10
8.0cb0.20
7.7cc0.10
6.8cd0.10
6.3ce0.15
3.9 cf0.15R
159
3.0 kGy +8l

cinnamaldehyde
Introduction
Summary
Summary
The consumption of fresh (unpasteurized) fruits and
vegetables juices has increased in recent years due to the
characteristics of freshness, high vitamin content, low calorie
contribution and an active promotion of fruits and vegetables
and their derivatives as important component of healthy diet for
immunocompromised patient.
Fresh fruit and vegetable juices are more perishable than
the raw fruits and vegetables from which they are processed and
usually have very short-life not exceeding 1-2 days at the best
refrigeration condition. This very short-life may be due to the
natural microbial flora (mainly yeasts and acid-love bacteria) of
raw fruits and vegetables as well as microbial contamination
during squeezing and packaging. Moreover, fresh fruits and
vegetables harbor some pathogens and a number of outbreaks of
foodborne illness have been reported due to the consumption of
fresh fruits and vegetables. Therefore, cold physical method has
to be used to extend the shelf-life of these fresh juices and to
eliminate contaminating pathogens without or with minimum
effect on their sensory qualities and nutrition. Thus, the main
objectives of the present study are:
1) Evaluate the microbiological quality of fresh pomegranate,
tomato and carrot juices.
2) Isolate the most predominant yeasts contaminating these
juices and investigate their sensitivity to gamma radiation
through determination of so - called D10 - value for each
isolated yeast.
3) Use of different doses of gamma radiation either alone or in
combination with other technologies to extend the shelf-life
of
160
Introduction
Summary
these juices and to inactivate the presence of pathogenic

bacteria.
4) Evaluate the sensorial and nutritional quality of the irradiated
juices during storage in order to identify the optimum
irradiation dose for each product and its shelf-life.
The results of this study can be summarized as follows:
Isolation and Identification of yeasts
Twenty yeast colonies were isolated from the juices under
investigation, pomegranate juice (7 colonies), tomato juice (8
colonies) and carrot juice (5 colonies). The identification
revealed that the 7 colonies from pomegranate juices were
Candida tropicalis (4 colonies) and Debaryomyces hansenii (3
colonies). Meanwhile, the 8 colonies from tomato juices were
Saccharomyces cerevisiae (5 colonies) and Trichosporon
pullulans (3 colonies). While, the 5 colonies from carrot juice
were Rhodotorula glutinis.
Radiation decimal reduction dose ( D10- value)
The effect of incremental doses of gamma irradiation
(0.0, 0.5, 1.0, 1.5, 2.0 and 2.5 kGy) on the viable count of the
isolated and identified yeasts, i.e. Rhodotorula glutinis, Candida
tropicalis, Trichosporon pullulans, Debaryomyces hansenii and
Saccharomyces cerevisiae were investigated. Generally, log
viable counts of tested microorganisms decreased with
increasing radiation dose but at different levels indicating that
these yeasts greatly differ in their sensitivity to gamma
irradiation. The D10-values of of, Candida tropicalis,
Rhodotorula glutinis, Saccharomyces cerevisia, Trichosporon
pullulans, Debaryomyces hansenii and in saline solution were
0.84, 0.92, 1.01, 1.25 and 1.29 kGy, respectively. Meanwhile
the D10-values of these yeasts
161
Introduction
Summary
in the juices were 0.98, 1.34, 1.34, 1.40 and 1.63 kGy,
respectively. It is obvious that Debaryomyces hansenii had the
highest D10-values, followed by Trichosporon pullulans
indicating that they had the highest radiation resistance in
comparison with the other studied yeasts. While, Candida
tropicalis had the lowest radiation resistance, i.e. had the highest
sensitivity to radiation.
Pomegranate juice
Effect of irradiation and refrigeration storage on the
microbial load, pathogenic bacteria, anthocyanin content, some
physiochemical properties and sensory evaluation of
pomegranate juice was investigated. It was found that the initial
microbial counts (total aerobic mesophilic bacteria, lactic acid
bacteria, molds and yeasts) of fresh pomegranate juice were
4.90, 4.10 and 3.96 log cfu/ml, respectively. The most probable
number of coliform bacteria was 75 MPN/ml, while Escherichia
coli, Enterococcus faecalis, Staphylococcus aureus and
Aeromonas hydrophila were below detectable level. Exposure
of fresh pomegranate juice to gamma radiation (1, 2 and 2.5
kGy) greatly reduced the initial microbial counts and completely
inactivated coliform bacteria.
During refrigeration storage of pomegranate juice at
4C1 there was a progressive increase in the viable counts of
aerobic mesophillic bacteria, molds and yeasts of irradiated and
non-irradiated pomegranate juice but the rate of increase was
lower in irradiated samples in comparison with unirradiated
ones.
All irradiation doses used (1, 2.0 and 2.5 kGy) decreased
anthocyanin content of pomegranate juice by 7.07, 14.00 and
15.45 %, respectively.
162
Introduction
Summary
The lowest irradiation dose used, i.e. 1 kGy had no effect

on the viscosity of pomegranate juice. Meanwhile, the other
irradiation doses used (2.0 and 2.5 kGy) reduced the viscosity
by 25 and 50 %, respectively. Irradiation at different doses used
had no effect on pH value of pomegranate juice.
The results of sensory panelists scores showed no
significant effect of irradiation at 1 kGy on the sensory
properties (color, odor, taste and overall acceptability).
Meanwhile, irradiation dose at 2.0 and 2.5 kGy reduced the
sensory panelist scores. From the view point of sensory
characteristics, panelists rejected unirradiated (control)
pomegranate juice samples after 7 days of refrigeration storage
because of off-odor and color changes. The same panelists
rejected pomegranate samples receiving 2.0 kGy and 2.5 kGy
after 14 days of refrigeration storage.
Tomato juice
microbial load, pathogenic bacteria, ascorbic acid content,
lycopene content, some physiochemical properties and sensory
evaluation of tomato juice was investigated. It was found that
the total log counts of aerobic mesophillic bacteria, lactic acid
bacteria, yeasts and molds of fresh tomato juice were 3.98, 3.67
and 3.57 cfu/ml, respectively. The most probable number of
coliform bacteria was 240 MPN/ml, while Escherichia coli,
Enterococcus faecalis, Staphylococcus aureus were below
detectable level. Irradiation of fresh tomato juice at different
doses (1.5, 3.0 and 4.5 kGy) caused a significant decrease in all
the initial microbial counts and completely inactivated coliform
bacteria.
163
Introduction
Summary
During refrigeration storage of tomato juice at 4C1 the

viable counts of aerobic mesophillic bacteria, lactic acid bacteria
and molds and yeasts in the unirradiated tomato juice were
progressively increased reaching 7.94, 6.96 and 7.07 log cfu/ml,
respectively after 10 days of storage. However, these microbial
counts in tomato juice receiving 1.5, 3.0 and 4.5 kGy reached
almost similar counts but after 15, 20 and 25 days.
All irradiation doses used (1.5, 3.0 and 4.5 kGy)
decreased ascorbic acid content of tomato juice by 15.35, 58.22
and 67.37 %, respectively. While the same irradiation doses
didnt affect the lycopene content in tomato juice.
Irradiation at (1.5, 3.0 and 4.5 kGy) lowered the viscosity
of tomato juice by 17.39, 39.13 and 47.82 %, respectively.
Irradiation at different doses used had no effect on pH value of
tomato juice.
The results of sensory evaluation revealed no significant
effect of irradiation at 1.5 and 3.0 kGy on the sensory properties
(color, odor, taste and overall acceptability). Meanwhile,
irradiation dose at 4.5 kGy didnt affect the color of the tomato
juice but affect the odor, taste and overall acceptability of
tomato juice samples. From the view point of sensory
characteristics, panelists rejected unirradiated (control) tomato
juice samples after 10 days of refrigeration storage because of
off odor, color changes and bad overall acceptability. The same
panelists rejected tomato samples receiving 1.5, 3.0 and 4.5 kGy
after 15, 20 and 25 days of storage at 4C1, respectively.
Carrot juice
microbial load, pathogenic bacteria, ascorbic acid content, total
164
Introduction
Summary
carotenoids content, some physiochemical properties and

sensory evaluation of carrot juice was investigated. It was found
that the initial microbial counts (aerobic mesophillic bacteria,
lactic acid bacteria, molds and yeasts) of fresh carrot juice were
4.98, 4.76 and 3.25 log cfu/ml, respectively. The most probable
number of coliform bacteria was (460 MPN / ml) E. coli (21
MPN/ml) and Enterococcus faecalis (3.5 x 10 2 cfu/ml). In
contrast Staphylococcus aureus and Aeromonas hydrophila
were not detected in unirradiated carrot juice. Gamma
irradiation of fresh carrot juice at 1.5, 3.0 and 4.0 kGy,
significantly reduced the microbial contamination level with a
dose-dependent tendency. A dose of 1.0 kGy or above
effectively reduced the coliform and E.coli population to below
the detectable level, while, irradiation doses of 3.0 and 4.0 kGy
reduced the counts of Enterococcus faecalis to below the
detectable level.
During refrigeration storage of tomato juice at 4C1,
there was a progressive increase in the viable counts of aerobic
mesophillic bacteria, lactic acid bacteria and molds and yeasts
of irradiated and non-irradiated carrot juice but the rate of
increase was lower in irradiated samples in comparison with
unirradiated ones indicating that irradiation retarded microbial
growth on storage. Total coliform and E.coli found in fresh
carrot juice decreased from 460 and 21 MPN/ml to 150 and < 3
MPN/ml, respectively at day 3. Meanwhile, Enterococcus
faecalis increased reaching 7.6 x 10 3 cfu/ml. Coliform bacteria,
E.coli and Enterococcus faecalis counts in all irradiated carrot
juice samples were below detectable level throughout the
refrigeration storage.
Gamma irradiation at dose of 1.5 kGy didnt show
changes in the ascorbic acid content, while doses of 3.0 and 4.0
kGy resulted in 25.79% and 55.11% loss of total ascorbic acid
content,
165
Introduction
Summary
respectively. While the same irradiation doses didnt seem to

decrease the total carotenoids content.
All irradiation doses used 1.5, 3.0 and 4.0 kGy had no
effect on the viscosity and pH value of carrot juice.
The result of sensory characteristics scores showed no
significant effect of irradiation at 1.5 and 3.0 kGy. However,
irradiation doses of 4.0 kGy affected the sensory characteristics
of carrot juice. Panelists rejected unirradiated (control) carrot
juice samples after 3 days of refrigeration storage because of
off-odor and very bad taste. The panelists rejected carrot juice
sample receiving 1.5 kGy after 5 days of refrigeration storage.
Meanwhile, carrot juice samples irradiated at 3.0 kGy and 4.0
kGy were sensorial rejected after 10 days of refrigerated
storage.
Combination treatment of irradiation and cinnamaldehyde
Effect of irradiation in combination with
cinnamaldehyde on the microbial load, pathogenic bacteria,
some physiochemical properties and sensory evaluation of
carrot juice was investigated. It was found that the initial
microbial counts of aerobic mesophillic bacteria, lactic acid
bacteria and molds and yeasts of fresh carrot juice was 4.25,
2.20 and 3.15 log cfu/ml. Addition of cinnamaldehyde alone at
concentration of 8l/100 ml reduced the counts of these
microorganisms by 1.01, 0.23 and 0.5 log cycles, respectively.
Meanwhile, addition of cinnamaldehyde at 8l/100 ml
concentration followed by irradiation at 3.0 kGy reduced all
microbial counts under investigation to below detectable level
(<10 cfu/ml).
During refrigeration storage, It was evident that all
microbial populations significantly increased as storage period
increase but the rate of increase was lower in the sample contain
166
Introduction
Summary
8l/100 ml cinnamaldehyde and was more lower in the sample

contain 8l/100 ml cinnamaldehyde and irradiation at 3.0 kGy.
Combination treatments of cinnamaldehyde and irradiation had
no significant effect either on viscosity or pH value of fresh
carrot juice.
The results of sensory evaluation scores showed no
significant effect on color, odor, taste and overall acceptability
of juices as a result of adding cinnamaldehyde alone or in
combination with irradiation at 3 kGy. During storage, the
sensory scores significantly decreased in all the samples but at
different rate. The rate of decrease in carrot juice containing
8l/100 ml cinnamaldehyde and irradiated at 3.0 kGy was the
lowest in comparsion with other treatments. The panelists
rejected carrot juice samples (control) at day 3 due to the offodor and bad taste. Carrot juice samples containing
cinnamaldehyde were organoleptically rejected after 7 day of
storage at 4C1. Combination treatment of cinnamaldehyde
and irradiation at 3.0 kGy extended the shelf-life of carrot juice
to 21 days from the view point of organoleptic quality.
Conculsion
A large part of the world's fresh fruits and vegetables are
processed into juices and other products. Gamma irradiation
alone or in combination with other techniques is used in this
study to preserve fresh pomegranate, tomato and carrot juices.
The conclusion drawn from the results obtained in this study
are:
1- Fresh pomegranate, tomato and carrot juices prepared
under the laboratory condition had generally high
microbial counts. All studied fresh juices contained
coliform bacteria;
167
Introduction
Summary
however, fresh carrot juice in addition to coliform

bacteria contained E.coli and Enterococcus faecalis.
2- Five yeast species have been isolated from these juices;
they are Rhodotorula glutinis, Candida tropicalis,
Trichosporon pullulans, Debaryomyces hansenii and
Saccharomyces cerevisiae.
3- The D10-value of these yeasts ranged from 0.98 to 1.63
kGy in the juice from which they were isolated.
Debaryomyces hansenii had the highest radiation
resistant (D10-value of 1.63 kGy) while Candida
tropicalis had the lowest radiation resistant (D10-value
of 0.98 kGy).
4- The lowest irradiation doses used (1- 1.5 kGy) were
sufficient to ensure the microbiological safety of these
fresh juices. Since they were very effective in
elimination of coliform bacteria and E.coli
contaminating fresh juices.
5- Irradiation dose of 2.5 kGy was the optimum dose for
preservation of fresh pomegranate juice. It extended the
shelf-life of fresh pomegranate juice to 14 days at
refrigeration temperature (4C1) with minimal
changes in its anthocyanin content and sensory
properties.
6- Irradiation dose of 3.0 kGy was effective in controlling
the microbial flora of tomato juice and extended its
shelf-life to 15 days at 4C1 against only 5 days for
unirradiated control without significant changes in
lycopene content.
168
Introduction
Summary
7- Irradiation dose of 3.0 kGy was also efficient in

reducing, to great extend, the microbial load of fresh
carrot juice and at the same time had no effect on
sensory characteristics or total carotenoid content, it
extend the shelf-life of fresh carrot juice to only 7 days
at 4C1, while the unirradiated (control) samples
organoleptically rejected at day 3.
8- Irradiation dose of 3.0 kGy in combination with
cinnamaldehyde (8l/100 ml) could extend the shelflife of fresh carrot juice to 21 days at 4C1 without
adverse effect on its sensory and nutritional quality.
169
Introduction
References
References
Abd El-Aziz, A.B. (2002): Effect of radiation on single cell
protein production, M.Sc. Thesis, Fac. Sci. Ain Shams
Univ., Cairo, Egypt.
Abd El-Aziz, A.B. and Abd El-Khalek, H.H. (2010): Non
chemical preservation of orange juice by natural
antimicrobial agents and gamma irradiation. Isotope.
.Rad.Res., 42,4(1),1301-1320.
Abu El-Nour, S.A. (2007): Microbiology and Radiobiological
studies on the hygienic quality of minimally processed
food, P.H.D. Thesis, Fac. Sci. Mansoura Univ., Cairo,
Egypt.
Abdellaoui, S.; Lacroix, M.; Jobin, M.; Boubekri, C. and
Gagnon, M. (1995): Effect of gamma irradiation
combined with hot water treatment on the physicochemical, nutritional and organoleptic qualities of
clementines. Science des Aliments, An. Int. J Food
Sci.Technol., 15, 217-235.
Adak, G.K.; Meakins, S.M.; Yip, H.; Lopman, B.A. and
OBrien, S.J. (2005): Disease risks from foods, England
and Wales, 19962000. Emerg. Infect. Dis., 11, 365372.
Adeil Pietranera, M.S.; Narvaiz, P.; Horak, C. and
Kairiyama, E. (2003): Irradiated icecreams for immunosuppressed patients. Radiat. Phys. Chem., 66, 357365.
Aker, S.N. and Cheney, C.L. (1983): The use of sterile and
low microbial diets in ultra-isolation environments.
JPEN., 7 (4), 390-397.
170
Introduction
References
Akhtar, M.H. and Bryan, M. (2008): Extraction and

quantification of major carotenoids in processed foods
and supplements by liquid chromatography. Food Chem.,
111, 255261.
Al. Maiman, S.A. and Dilshad, A. (2002): Changes in physical
and chemical properties during pomegranate (Puncia
granatum) fruit maturation. Food Chem., 76, 437-441.
Alighourchi, H.; Barzegar, M. and Abbasi, S. (2008): Effect
of gamma irradiation on the stability of anthocyanins and
shelf-life of various pomegranate juices. Food Chem.,
110(4), 1036-1040.
Alvarado, J.D. and Romero, C.H. (1989): Physical properties
of fruits I II. Density and viscosity of juices as functions
of soluble solids content and temperature. Latin American Appl. Res., 19, 1521.
Anastasakis, M.; Lindamood, J.B.; Chism, G.W. and
Hansen, P.M.T.(1987): Enzymatic hydrolysis of carrot
for extraction of a cloud-stable juice. Food Hydrocolloids.
1 (3), 247261.
Anon (1999): Safe tables our priority: technical meetings on
juice safety (online).
Http://stop-usa.org/news/priorcom/11999com.html
Anonymous (1990): Vitamins and the immune response.
Medical update, Number 4, October 1990. Isando, South
Africa:Vitamin Information Centre.
Anonymous (2000 a): Food irradiation: Available research
indicates that benefits outweigh risks, U.S. General
Accounting Office, GAO/RCED -00-217.
171
Introduction
References
Anonymous (2000 b): Irradiation in the production, processing

and handling of food, Code of Federal Regulations,
21CFR179.
AOAC (2000): Official methods of analysis. Association of
Official Analytical Chemists.16th ed., Maryland, USA.
APHA (1992): Standard methods for the examination of dairy
products.AmericanPublic
Health
Association,
Washington, D.C.
Arias, M.T.G.; Pontes, E.A.; Fernandez, M.C.G. and Muniz,
F.J. S. (2003): Influence of slow and quick defrosting on
protein quality. J. Sci. Food.Agric., 83,602608.
Arts-Hernandez, F.; Roblesa, P.A.; Gomez, P.A.; TomasCallejasa, A. and Artsa, F. (2010): Low UV-C
illumination for keeping overall quality of fresh-cut
watermelon, Postharv. Bio.Technol., 55 ,114120.
Aune, E.J. (2003): Superchilling of foodstuff. A review, 21th
Congress ICR, August17 22,2003,Washington,DC,
<http://www.icr2003.org>.
Aviram, M.; Dornfeld, L. and Rosenblat, M. (2000):
Pomegranate juice consumption reduces oxidative stress,
atherogenic modifications of LDL, and platelet
aggregation: studies in humans and in atherosclerotic
apolipoprotein
E-deficient
mice.Am.J.Clin.Nutr.,
71,106276.
Ayiek, H.; Aydogan, H. and Kuukkaraaslan, A. (2004):
Assessment of the bacterial contamination on hands of
hospital food workers. Food Control., 15, 253-259.
172
Introduction
References
Ayed, N.; Yu, H. L. and Lacroix, M. (1999): Improvement of

anthocyanin yield and shelf-life extension of grape
pomace by gamma irradiation. Food Res. Int., 32, 539543.
Aymerich, T.; Picouet, P. A. and Monfort, J. M. (2008):
Decontamination technologies for meat products. Meat
Sci., 78, 114129.
Aziz, N.H. and Moussa, L.A.A. (2002): Influence of gammaradiation on mycotoxin producing moulds and
mycotoxins in fruits. Food Control., 13, 281288.
Azzouz, M. A. and Bullerman, L. B. (1982): Comparative
antimycotic effects of selected herbs, spices, plant
components and commercial fungal agents. J. Food Sci.,
45, 1298-301.
Badosa, E.; Trias, R.; Pars, D.; Pla, M. and Montesinos,
E.(2008): Microbiological quality of fresh fruit and
vegetable products in Catalonia (Spain) using normalised
plate counting methods and real time polymerase chain
reaction (QPCR). J. Sci. Food Agri., 88, 605611.
Bakowska, A.; Kucharska, A. Z. and Oszmianski, J. (2003):
The effects of heating, UV irradiation, and storage on
stability of the anthocyanin polyphenol co-pigment
complex. Food Chem., 81(3), 349-355.
Barbosa-Canovas,G.V.;Gongora-Nieto,
M.M.;
Pothakamury,
U. R. and Swanson, B.G. (1998):
Water Activity in Foods: Fundamentals and Applications
ISBN: 978-0-8138-2408-6, September 2007, WileyBlackwell.
173
Introduction
References
Barkai-Golan, R. (1992): Suppressing of Postharvest

Pathogens of Fresh Fruits and Vegetables by Ionizing
Radiation. In: I. Rosenthal (ed.) Electromagnetic
Radiation in Food Science,Springer-Verlag, Berlin,
Heidelberg, pp. 155-193.
Barnett, J.A.;Payne, R.W. and Yarrow, D. (1990): Yeasts:
Characteristics and Identification. 2nd Edn. Cambridge
University press, Cambridge.
Barnett, J.A.; Payne, R.W. and Yarrow, D. (2000): Yeasts:
Characteristics and Identification. 3rd Edn. Cambridge
University press, Cambridge.
Baskaran, S.A.; Amalaradjou, M.A.R.; Hoagland, T. and
Venkitanarayanan, K. (2010): Inactivation of E. coli
O157:H7 in apple juice and apple cider by transcinnamaldehyde. Int.J.Food Microbiol., 141,126129.
Bawa, A.S. and Saini, P.S. (1987): Effect of method of
preservation on the storage quality of carrot juice. Indian
Food Packer., 41 (1), 4246.
Beech, F.W. and Davenport, R.R. (1970): The role of yeasts
in cider making. In: Rose, A.H., Harrison, J.S. (Eds.), The
Yeasts. Academic Press, London, pp. 73146.
Ben, A.M. (1999): Effect of freezing and microbial growth on
myoglobin derivates of beef. Food Chem., 147, 4093.
Beuchat, L.R. (1996): Pathogenic microorganisms associated
with fresh produce. J. Food Prot., 59(2), 204-216.
Beuchat, L.R. (1998): Surface decontamination of fruits and
vegetables eaten raw: A review. Food Safety Unit World
Health Organization, WHO/FSF/98.2.
174
Introduction
References
Bibbington, A.; Wilson, P. and Jones, M. (1993): Audit of

nutritional advice given to bone marrow transplant
patients in the United Kingdom and Eire., Clin. Nutr., 12,
230-235.
Bodey, G.P.; Buckley, M.; Sathe, Y.S. and Freireich, E.J.
(1966): Quantitative relationships between circulating
leukocytes and infection in patients with acute leukaemia.
Ann. Internal.Medicine. 64 (2), 328340.
Bognar, A. (1998): Comparative study of frying to other
cooking techniques influence on the nutritive value.
Grasas Aceites. 49, 250260.
Boskovic, M. (1979): Fate of lycopene in dehydrated tomato
products: carotenoids isomerization in food systems. J.
Food. Sci., 44, 8486.
Brand, S.; Fardel, G.; Flandrois, J.P.; Rosso, L. and
Tomassone, R. (1999): A model describing the
relationship between re-growth lag time and mild
temperature increase for Listeria monocytogenes. Int. J.
Food Microbiol., 46 (3), 251261.
Breslin, P. and Spector, A.C. (2008): Mammalian taste
perception. Current Biology, 18, R148-R155.
Britton, G. (1992): Carotenoids. In Natural Foods Colorants,
ed. G. F. Hendry, pp. 141-148. G. F. Blackie, New York.
Buchanan, R.L.; Edelson, S.G.; Snpes, K. and Boyd, G.
(1998): Inactivation of E. coli O157:H7 in apple juice by
irradiation. Appl. Environ. Microbiol. 64 (11), 4533
4535.
Bullerman, L.B. (1977): Inhibition of aflatoxin production by
cinnamon. J. Food Sci., 39, 1163-5.
175
Introduction
References
Burt, S. (2004): Essential oils: Their antibacterial properties

and potential applications in foods. A review. Int.J.Food
Microbiol., 94: 223253.
Butterweck, J.S. (1995): Sterile diets for the immunocompromised: is there a need? Radiat. Phys. Chem. 46,
(4-6), 601-604.
Byun, M.W.; Kim, D.H.; Yook, H.S.; Cha, B.S. and Kim,
J.O. (2001): Changes in microbiological and general
qualities in gamma irradiated Doenjang(fermented
soybean paste ). Food sci. biotechnol., 10,7-11.
Cam, M.; Hisil, Y. and Durmaz, G. (2009): Classification of
eight pomegranate juices based on antioxidant capacity
measured by four methods. Food Chem., 112,721-726.
Chachin, K. and Ogata, K. (1969): Changes of chemical
constituents and quality in some juice irradiated with the
sterilizing dose level of gamma rays. Food Irradiat., 4(1),
85-90.
Chanock, S.J. and Pizzo, P.A. (1995): Infection prevention
strategies for children with cancer and AIDS: contrasting
dilemmas, J.Hosp.Infect., 30 (Supplement), 197-208.
Chaudry, M.S.; Bibi, N.; Khan, M.; Khan, M.;
Badshah, A.; and Qureshi, M.J. ( 2004 ): Irradiation
treatment of minimally processed carrots for ensuring
microbiological safety. Radiat. Phys. Chem., 71,169-173.
Chervin, C. and Biosseau, P. (1994): Quality maintenance of
ready-to-eat shredded carrots by gamma radiation. J.
Food Sci. 59, 359365.
176
Introduction
References
Chiasson, F.; Borsa, J.; Ouattara, B. and Lacroix, M. (2004):

Radiosensitization of Escherichia coli and Salmonella
typhi in ground beef. J. Food Prot., 67, 1571162.
Christensen, E.A. (1970): Radiation resistance of bacteria and
the microbiological control of irradiated medical
products, sterilization and preservation of biological
tissues by ionizing radiation. IAEA, Vienna.
Codex stan 49-1981. Codex standard for tomato juice preserved
exclusively by physical means (world-wide standard).
Collins, C.H.; Lyne, P.M. and Grange, J.M. (1989): Collins
and Lynes Microbiological Methods sixth ed.
Butterworths, London.
Conner, D.E. and Beuchat, L.R. (1984): Effects of essential
oils from plants on growth of food spoilage yeasts. J.
Food Sci., 49, 429-224.
Considine, K.M.; Kelly, A.L.; Fitzgerald, G.F.; Hill, C. and
Sleator, R.D. (2008): High-pressure processing-effects
on microbial food safety and food quality. FEMS
Microbiology Letters, 281(1), 1-9.
Cook, N.C. and Samman, S. (1996): Flavonoids chemistry
metabolism, cardioprotective effects, and dietary sources.
J. Nutr. Biochem., 7 ,66-76.
Cook, K.A.; Dobbs, T.E.; Hlady, W.G.; Wells, J.G.; Barrett,
T.J.; Puhr, N.D.; Lancette, G.A.; Bodager, D.W.;
Toth, B.L.; Genese, C.A.; Highsmith, A.K.; Pilot,
K.E.; Finelli, L. and Swerdlow, D.L. (1998): Outbreak
of Salmonella serotype Hartford infection associated with
un-pasteurized orange juice. JAMA., 280 (17), 1504
1509.
177
Introduction
References
Crelier, S.; Robert, M. C.; Claude, J. and Juillerat, M. A.

(2001): Tomato (Lycopersicon esculentum) pectin
methyl-esterase
and
polygalacturonase
behaviors
regarding heat-and pressure-induced inactivation. J.
Agric.Food Chem., 49(11), 55665575.
Cuvielier, M.E.; Berset, C. and Richard, H. (1994): Antioxidant constituents in sage (Salvia officinalis). J. Agric.
Food Chem., 42, 655-669.
Daw, M.A. and Falkiner, F.R. (1994): Prevention of Gramnegative infections in neutropenic cancer patients. Saudi
Medical Journal., 15 (3), 196203.
DeMille, D.; Deming, P.; Lupinacci, P.; and Linda, A. and
Jacobs, L.A. (2006): The effect of the Neutropenic diet in
the outpatient setting: A pilot study. Onco. Nur. For., 33:
337
Dezenhall, A.; Curry-Bartley, K.; Blackburn, S.A.; De
Lamerens , S. and Khan, A.R.(1987): Food and
nutrition services in bone marrow transplant centres. J.
Am. Diet Assoc., 87 (10), 1351-1353.
Diehl, J.F. (1995): Safety of Irradiated Foods (2nd edition).
Marcel Dekker Inc., 1995. ISBN 0 82479 344 7. 464 pp.
DiMascio, P.; Aaiser, S. and Sies, H. (1989): Lycopene as the
most effective biological carotenoid singlet oxygen
quencher. Arch. Biochem. Biophys., 274, 532-538.
Dorais, M.; Ehret, D.L. and Papadopoulos, A.P. (2008):
Tomato (Solanum lycopersicum) health components: from
the seed to the consumer. Phyto.Chem.Rev.,7, 231 250.
Doyle, M.P. (1990): Fruit and vegetable safety microbiological
considerations. Hort.sci., 25, 14781481.
178
Introduction
References
Dris, R and Jain, S.M. (Eds.) (2004): Production Practices and

Quality Assessment of Food Crops,Vol. 4,Postharvest
Treatment and Technology, pp. 261295. 2004 Kluwer
Academic Publishers. Printed in the Netherlands.
Du, C.T.; Wang, P.L. and Francis, F.J. (1975): Anthocyanins
of pomegranate ( Punica granatum). J. Food Sci. 40,
417418.
Dumas, Y.; Dadomo, M.; Di Lucca, G. and Grolier, P.
(2003): Effects of environmental factors and agricultural
techniques on antioxidant content of tomatoes. J. Sci.
Food Agric., 83, 369382.
Duodua, K.G.; Minnaara, A. and Taylor, J.R.N. (1999):
Effect of cooking and irradiation on the labile vitamins
and antinutrient content of traditional African sorghum
porridge and spinach relish, Food Chem., 66, 21-27.
Erkan, M.; Wang, S.Y. and Wang, C.Y. (2008): Effect of UV
treatment on antioxidant capacity, antioxidant enzyme
activity and decay in strawberry fruit. Postharv. Biol.
Technol., 48, 163-171.
Espina, L.; Somolinos, M.; Lorn, S.; Conchello, P.; Garca,
D. and Pagn, R. (2011): Chemical composition of
commercial citrus fruit essential oils and evaluation of
their antimicrobial activity acting alone or in combined
processes. Food Control., 22, 896-902.
Fan, X.; Niemira, B.A. and Sokorai, K.J.B. (2003a): Use of
ionizing radiation to improve sensory and microbial
quality of fresh-cut green onion leaves. J Food Sci., 68,
14781483.
Fan, X.; Niemira, B. and Skorai, K.J.B., (2003b): Sensorial,
nutritional and microbiological quality of fresh cilantro
179
Introduction
References
leaves as influenced by ionizing radiation and storage.

Food Res.Int., 36,649-760.
Farkas, J. (1990): Combination of irradiation with mild heat
treatment. Food Control., 1, 223-229.
Farkas, J.; Saray, T.; Mohacsi-Farkas, C.; Horti, C. and
Andrassy, E. (1997): Effects of low-dose gamma
irradiation on shelf life and microbiological safety of
precut/prepared vegetables. Adv.Food Sci. 19, 111 119.
Farkas, J. and Mohacsi-Farkas, C.(2010): History and future
of food irradiation. Trends in Food Sci. and Technol.,
22(2-3), 121-6
Finberg, R.W. and Talcott, J.A.(1999): Fever and
neutropenia- how to use a new treatment strategy. New
England Journal of Medicine., 341 (5), 362363.
Fishman, M. and Mrozek-Orlowski, M. (Eds.). (1999):
Cancer chemotherapy guidelines and recommendations
for practice ( 2nd ed. ). Pittsburgh, PA: Oncology Nursing
Press, Inc.
Foley, D.M.; Dufour, A.; Rodriguez, L.; Caporaso, F. and
Prakash, A. (2002): Reduction of E.coli 0157:H7 in
shredded iceberg lettuce by chlorination and gamma
irradiation. Radiat. Phys. Chem., 63, 391396.
Frankmann, C.F.; Beck, J. and homburg, R.K. (1984):
Feeding in protected environments. In: Rose J C, ed.
Handbook for health care food service management, 241250.
Fried, E.J. and Nestle, M. (2002): The growing Political
Movement against Soft Drinks in Schools. J. Am. Med.
Assoc., 288(17), 2181.
180
Introduction
References
Gagnon, M.; Julien, J.P. and Riel, R.R. (1968): Irradition of

apple juice. I. Effects of gamma rays on the rate of
filteration and the viscosity. J. Candian Institute of Food
technology, 1 (4), 117-122.
Gallart-Jornet, L.; Rustad, T.; Barat, J.M; Fito, P. and
Escriche, I. (2007): Effect of superchilled storage on the
freshness and salting behavior of Atlantic salmon
(Salmo salar) fillets. Food Chem. 103 (4), 12681281.
Garca-Closas, R.; Berenguer, A.; Jos Tormo, M.; Jos
Snchez, M.; Quirs, J.R.; Navarro, C.; Arnaud, R.;
Dorronsoro, M.; Dolores Chirlaque, M.; Barricarte,
A.; Ardanaz, E.; Amiano, P.; Martinez, C.; Agudo, A.
and Gonzlez, C.A. (2004): Dietary sources of vitamin
C, vitamin E and specific carotenoids in Spain. Br. J.
Nutr., 91, 1005-1011.
Gardner, L.K. and Lawrence, G.D.(1993): Benzene
production from decarboxylation of benzoic acid in the
presence of ascorbic acid and a transition-metal catalyst.
J.Agric. Food Chem., 41,693-695.
Gerster, H. (1997). The potential role of lycopene for human
health. J. Am.College Nutr., 16, 109-126.
Ghaffari, M.A. and Ghiasvand, T. (2006): Effect of lycopene
on formation of low density lipoprotein-copper complex
in copper catalyzed peroxidation of low density
lipoprotein, as in vitro experiment, Iranian. Biomedic. J.,
10 (4), 191-196.
Ghosh, M.; Ganguli, A. and Mudgil, S. (2004): Microbiological quality of carrots used for preparation of fresh
squeezed street vended carrot juices in India. Food.
Agric.Environ., 2(2), 143-145.
181
Introduction
References
Gil, M.I.; Tomas-Barberan, F.A.; Hess-Pierce, B.; Holcroft,

D.M. and Kader, A.A. (2000): Antioxidant activity of
pomegranate juice and its relationship with phenolic
composition and processing. J. Agric. Food Chem., 48,
4581 9.
Giovannucci, E. (1999): Tomatoes, tomato-based products,
lycopene, and cancer: review of the epidemiologic
literature. J. Natl. Cancer. Inst., 91, 317-331.
Giusti, M.M. and Wrolstad, R.E. (2001): Characterization and
measurement of anthocyanin by UV- visible
spectroscopy. In : Wrolstad, R.E, Schwartz, S.J. (Eds.),
Current protocols in food analytical chemistry. John wiley
and sons, New York, pp. 1-13.
Gonzalez-Aguilar, G. A.; Villegas-Ochoa, M. A.; MartnezTellez, M. A.; Gardea, A. A. and Ayala-Zavala, J. F.
(2007): Improving antioxidant capacity of fresh-cut
mangoes treated with UV-C. J. Food Sci.,72, 197-202.
Goto, K.; Mochida, K.; Asahara, M.; Suzuki, M.; Kasai, H.
and Yokota, A. (2003): Alicyclobacillus pomorum sp.
nov., a novel thermo-acidophilic, endospore-forming
bacterium that does not possess-alicyclic fatty acids, and
emended description of the genus Alicyclobacillus. Int. J.
Syst. Evol. Microbiol. 53, 1537-1544.
Goularte, I.; Martins, C.G.; Morales-Aizpurua, I.C.; Destro,
M.T.; Franco, B.D.G.M.; Viozeu, D.M.; Hutzler, B.W.
and Landgraf, M. (2004): Combination of minimal
processing and irradiation to improve the microbiological
safety of lettuce. Rad. Phys. Chem., 71,157-161.
Graham, W.D. and Stevenson, M.H. (1997): Effect of
irradiation on vitamin C content of strawberries and
182
Introduction
References
potatoes in combination with storage and with further

cooking in Potatoes. J. Sci. Food Agric., 75, 371-377.
Grinbaum, A.; Ashkenazi, I.; Treister, G.; GoldschmiedRequven, A. and Block, C.S. (1994): Exploding bottles:
eye injury due to yeast fermentation of an uncarbonated
soft drink. Br. J. Ophthalmol., 78 (11), 883.
Groenewald, W.H.; Gouws, P.A. and Witthuhn, R.C. (2009):
Isolation, identification and typification of Alicyclobacillus acidoterrestris and Alicyclobacillus acidocaldarius strains from orchard soil and the fruit
processing environment in South Africa. Food Microbiol.,
26, 71-76.
Gu, L.; Kelm, M.; Hammerstone, J.F.; Beecher, G.; Holden,
J. and Haytowitz, D. (2004): Concentrations of proanthocyanidins in common foods andestimations of
normal consumption. J. Nutr., 134, 6137.
Guerrero-Beltrn, J.A. and Barbosa-Cnovas, G.V. (2005):
Reduction of Saccharomyces cerevisiae, E.coli and
Listeria innocua in apple juice by ultraviolet light. J. of
Food Process Engineering., 28, 437452.
Hagenmaier, R.D. and Baker, R.A. (1998): Microbial
population of shredded carrot in modified atmosphere
packaging as related to irradiation treatment. J. Food Sci.,
63, 162164.
Hajare, S.N.; Saraj. S.D.; Dhokane, V.S.; Shashidhar, R.
and Bandekar, J.R. (2007): Effect of radiation
processing on nutritional and sensory quality of
minimally processed green gram and garden pea sprouts.
Radiat.Phys Chem., 76, 16421649.
183
Introduction
References
Hamilton-Miller, J.M. and Shah, S.(2001): Identity and

antibiotic susceptibility of enterobacterial flora of salad
vegetables. Int. J. Antimicrob. Agents., 18, 8183.
Hammad, A.A.I.; Mongy, T.M.; Abu-Shady, M.R. and
Taha, S.M. (1995): Microbial changes in strawberries
treated with gamma irradiation to improve their quality.
Egypt. J.Food. Sci., 23, 117-132.
Hammad, A.A.; Gebreel, H.M.; Taha, S.M. and Amal, S.M.
(2010): Effect of gamma irradiation and refrigeration
storage on microbial, sensorial and chemical quality of
fresh-cut lettuce. Egypt. J.Rad.Sci., Applic.23, 143-157.
Hanotel, L.; Fleuriet, A. and Boisseau, P. (1995):
Biochemical changes involved in browning of gammairradiated cut wit loof chicory. Postharvest Bio.Technol.,
5, 19921.
Harris, L.J.; Farber, J.N.; Beuchat, L.R.; Parish, M.E.;
Suslow, T.V. and Garrett, E. H. (2003): Outbreaks
associated with fresh produce: Incidence, growth, and
survival of pathogens in fresh and fresh-cut produce. In
comprehensive reviews in food science and food safety
(pp.78141).Iowa:BlackwellPublishingProfessional.
<http://www.ift.org> Accessed 20.12.2004.
Hayashi, T. and Todoriki, S. (1996): Detection of irradiated
peppers by viscosity measurement at extremely high pH.
Radiat.Phys.Chem., 48, 101-104.
Hayes, W.A.; Smith, P.G. and Morris, A.E.J. (1998): The
production and quality of tomato concentrates. Crit Rev
Food Sci Nutr., 38(7), 537564.
Head, K.A. (1998): Ascorbic acid in the prevention and
treatment of cancer, Alt. Med. Rev., 3, 174-186
184
Introduction
References
Hegazi, R.A. (1981): Radiation-induced changes in the

chemical constituents of strawberry fruit and derivative.
M.Sc. Thesis, Fac. Agric., Mansoura Univ.
Helander, I.M.; Alakomi, H.L.; Latva-Kala, K.; MattilaSandholm, T.; Pol, I.; Smid, E.J.; Gorris, L.G.M. and
von Wright, A. (1998). Characterization of the action of
selected essential oils components on Gram negative
bacteria. J. Agric. Food Chem., 46, 35903595.
Hell, K.; Gnonlonfin, B.G.J.; Kodjogbe, G.; Lamboni, Y.
and Abdourhamane, I.K. (2009): Mycoflora and
occurrence of aflatoxin in dried vegetables in Benin, Mali
and Togo, West Africa , Int.J. Food Microbiol., 135 , 99
104.
Hernandez, F.; Melgarejo, P.; Tomas-Barberan, F.A. and
Artes, F. (1999): Evolution of juice anthocyanin during
ripening of new selected pomegranate (Punica granatum)
clones. Eur. Food Res. Technol., 210, 3942.
Howard, L.R.; Miller, G.H.; Wagner, A.B.(1995): Microbiological, chemical and sensory changes in irradiated
Pica De Gallo. J. Food. Sci. 60(3), 461-464.
Hsu, K.C.; Tan, F.J. and Chi, H.Y (2008): Evaluation of
microbial inactivation and physiochemical properties of
pressurized tomato juice during refrigeration storage.
LWT. 41, 367375.
Huis int Veld, J.H.J. (1996): Microbial and biochemical
spoilage of foods: an overview. Int.J.Food.Microbiol.,
33(1-18).
185
Introduction
References
Hussain, P.R.; Meena, R.S.; Dar, M.A. and Wani, A.M.

(2008): Studies on enhancing the keeping quality of peach
(Prunus persica Bausch) Cv.Elberta by gammairradiation. Rad. Phys. Chem.; 77, 473481
IARC (2003):Fruit and Vegetables: IARC Handbooks of
Cancer Prevention.International Association of Research
on Cancer. IARC Press, Lyon, France.
Ibrahim, O.I.S. (1996): Studies on the biochemical changes
induced by gamma rays in carrot plants. M.Sc. Thesis,
Fac. Agric., Ain Shams Univ.
ICGFI (1999): Facts about food irradiation: a series of fact
sheets from the International Consultative Group on Food
Irradiation. ICGFI, Vienna.
ICMSF (1978): International Commission on Microbiology
Specifications for food. Microorganisms in foods. 1. Their
significance and methods of enumeration, 2nd edn.
Toronto: University of Toronto Press.
IFT (1998): Institute of Food Technologies, Extending shelflife of refrigerated foods: Microbiological quality and
safety, 52(2), 57-62.
Ingram, M. (1975): Microbiology of foods pasturized by
ionizing radiation Technical report series IFIP-R 33.
International Project in the field of Food Irradiation,
Karlsruhe.
Ingram, M. and Roberts, T. A. (1980): Ionizing irradiation. In
microbial Ecology of foods vol.I : factors affecting life
and death of microorganisms. ICMSF (Ed.), Academic
Press, New York. pp. 46. Irradiation, 4(1), 8590.
186
Introduction
References
Jimenez, F.; Garro, L.; Rodrique, Z.E. and Zeledon, Z.

(2004) : Evaluation of the presence of bacteria in food
and environment of an oncological service of national
hospital, San Jose, Costa Rica. Arch. Latinoam. Nutr., 54,
303-307.
Jo, C.; Kim, H.J.; Kim, D.H.;Lee, W.K.; Ham, J.S. and
Byun, M.W.( 2007): Radio sensitivity of selected
pathogens in ice cream. Food Control, 18, 859865.
Josimovic, l. (1983): Study on some chemical changes in
irradiated pepper and parsley. Int. J. Appl. Radiat. Isot.,
34 (5), 787791.
Juszczak, L. and Fortuna, T. (2004): Effect of temperature
and soluble solid content on the viscosity of cherry juice
concentrate. International Agrophysics, 18, 1721.
Kamat, A.; Pingulkar, K.; Bhushan, B.; Gholap, A. and
Thomas, P. (2003): Potential application of low dose
gamma irradiation to improve the microbiological safety
of fresh coriander leaves. Food Control, 14, 529537.
Kandler, O. and Weiss, N. (1986): In: Bergey's Manual of
Systematic Bacteriology, P. H. A. Sneath, N. S. Mair, M.
E. Sharpe, J. G. Holt
Karel, M.; Fernema, 0. R. and Lund, D. B. (1975): Principles
of Food Science. Part II Physical Principles of Food
Preservation. (Ed. Owen, R.) Fernema. Marcel Dekker,
Inc. 270 Madison Avenue, New York, NY. 10016. pp. 33
- 36.
Kayacier, A. and Dogan, M. (2006): Rheological properties of
some gums-salep mixed solutions. J. Food Engin.,
72(3), 261265.
187
Introduction
References
Keijer, J.; Bunschotena, J.E.; Paloub, A. and FranssenHala, N.L.W.(2005): Beta-carotene and the application
of transcriptomics in riskbenefit evaluation of natural
dietary components, Biochimica et Biophysica Acta.,
1740, 139 146.
Keyser, M.; Muller, I. A.; Cilliers, F. P.; Nel, W. and
Gouws, P.A. (2008): Ultraviolet radiation as a nonthermal treatment for the inactivation of microorganisms
in fruit juice.Innovative Food Science and Emerging
Techno-logies, 9(3), 348-354.
Kilcast, D. (1994): Effect of irradiation on vitamins. Food
Chem., 49, 157164
Kilcast, D. and Subramaniam, P. (2000): Stability and shelf
life of food. Leather head Food Research Association.
<http://www.knovel.com/web/portal/browse/display?_EX
T_KNOVEL_DISPLAY_bookid=14>
Kim, K.H. and Yook, H.S. (2009): Effect of gamma irradiation
on quality of kiwi fruit (Actinidiadeliciosa var. deliciosa
cv.Hayward). Radiat. Phys.Chem., 78, 414421.
Kim, D.; Song, H.; Lim, S.; Yun, H. and Chung, J. (2007):
Effects of gamma irradiation on the radiation-resistant
bacteria and polyphenol oxidase activity in fresh kale
juice. Radiat. Phys. Chem.,76, 12131217.
Koc, A.N.; Silici, S.; Sariguze, F.M. and Sagdic, O. (2007):
Antifungal activity of propolis in four different fruit
juices. Food Technol. Biotechnol., 45 (1), 57-61.
Koseki, P.M.; Villavicencio, A.L.C.H.; Brito, M.S.; Nahme,
L.C.; Sebastiao, K.I. and Rela, P.R. (2002): Effects of
irradiation in medicinal and eatable herbs. Radiat.Phys.
Chem., 63, 681-684.
188
Introduction
References
Krinsky, N.I. and Johnson, E.J. (2005): Carotenoid actions

and their relation to health and disease. Mol.Aspects Med.,
26,459516.
Kumar, R. and Vaithiyanathan, S. (1990): Occurrence,
nutritional significance and
effect
on
animal
productivity of tannins in tree leaves. Animal Feed
Sci.Technol., 30, 21-38.
Kurtzman, C.P. and Fell, J. W. (2000): The yeasts,
ataxonomic study, 4th ed., Amsterdam: Elsevier.
Lacroix, M. and Gagnon, M (1993): Effect of gamma
irradiation with or without hot water dip and
transportation from Thailand to Canada on nutritional
qualities, ripening index and sensorial characteristics of
Thai mangoes (Nahng Glahng Wahn variety).
Radi.Phys.Chem., 42(1-3), 273-277.
Lacroix, M. and Ouattara, B. (2000): Combined industrial
processes with irradiation to assure innocuity and
preservation of food products- a review. Food Res.Inter.,
33, 719-724.
Lacroix, M.; Jobin, M.; Laterille, B.; Laponite, M. and
Gagnon, M. (1991) : Hot water immersion and
irradiation effect on mangoes keeping quality after air
shipment from Thailand and Canada. Microbial.Alim.
Nutr., 9, 155160.
Lacroix, M.; Turgis, M.; Borsa, J.; Millette, M.; Salmieri, S.;
Caillet, S. and Han, J. (2009): Applications of radiation
processing in combination with conventional treatments
to assure food safety: New development. Radiat.Physi.
Chem., 78, 1015-1017.
189
Introduction
References
Ladaniya, M.S.; Singh, S. and Wadhawan, A.K. (2003):

Response of Nagpur mandarin, Mosambi sweet orange
and Kagzi acid lime to gamma radiation. Radiat. Phys.
Chem., 67, 665-675.
Lafortune, R. and Lacroix, M. (2004): Combined effects of
gamma irradiation and modified atmosphere packaging on
shelf life extension of grated carrots (Daucus carota).
Radiat. Phys. Chem., 71, 7780.
Lalaguna, F. (2003): Physicochemical response of palmita-type
cheese to low-dose irradiation. J. Food Sci., 68, 2630.
Langley, P.(2000): Why a pomegranate? Br Med J., 321,1153
4.
Lawerence, C.E. (1971): Cellular radiobiology. William and
Sons. Ltd., London.
Lawless, H.T. and Heymann, H. (1999): Sensory evaluation of
food: Principles and practices. Gaithersburg: Aspen.
Lee, J.H.; Sung, T.H.; Lee, K.T.and Kim, M.R. (2004):
Effect of gamma irradiation on color , pungency , and
volatiles of Korean red pepper powder. J.Food Sci., 69,
585-590.
Lee, N.Y.; Jo, C.; Shin, D.H.; Kim, W.G. and Byun, M.W.
(2006): Effect of -irradiation on pathogens inoculated
into ready-to-use vegetables. Food Microbiol., 23, 649656.
190
Introduction
References
Lee, J.W.; Kim, J.K.; Srinivasan, P.; Choi, J.; Kim, J.H.;
Han, S.B.; Kim,D.J. and Byun, M.W. (2009): Effect of
gamma irradiation on microbial analysis,antioxidant
activity, sugar content and color of ready- to-use tamarind
juice during storage. LWT- Food Sci. Technol., 42, 101105.
Lir, S.; Manson, J.E. and Lee, I.M. (2000): Fruit and
vegetable intake and risk of cardio vascular disease.
Am.J.Clin. Nutr., 72, 922930.
Liu, R.H. (2004): Potential synergy of phytochemicals in
cancer prevention: mechanism of action. The Journal of
Nutrition (Supplementary Issue) 3479-3485.
Lopez-Rubira, V.; Conesa, A.; Allende, A. and Arte s, F.
(2005): Shelf life and overall quality of minimally
processed pomegranate arils modified atmosphere
packaged and treated with UV-C. Postharv. Bio.Technol.,
37, 174-185.
Loaharanu, P. (1996): Irradiation as a cold pasteurization
process of food. Veterinary Parasitology. 64, 71-82.
Lombrana, J.I. and Dias, J.M. (1985): Rheological and
chemical changes in stored carrot Juice. Canadian
Institute of Food Science and Technology Journal. 18 (3),
2132119.
Lu, Z.; Yu, Z.; Gao, X.; Lu, F. and Zhang, L. (2005):
Preservation effects of gamma irradiation on fresh-cut
celery. J.Food Engineering, 67,347-35.
Ludwig, D.S.; Eterson, K.E., and Gortmaker, S.L. (2001):
Relation between consumption of sugar-sweetened drinks
and childhood obesity: a prospective, observational
analysis. The Lancet. 357, 505508.
191
Introduction
References
Lugauskas, A.; Raudoniene, V. and Sveistyte, L. (2005):

Toxin producing micromycetes on imported products of
plant origin. An. Agri. Environ. Medicine., 12, 109118.
Lundgren, B.; Elvin, K. and Rothman, L.P. (1997):
Transmission of Pneumocytis carinii from patients to
hospital staff. Thorax ,52, 422-4.
Machado, T.; Pinto, A.; Pinto, M.; Leal, I.; Silva, M.;
Amaral, A.; Kuster, R. and Netto dosSantos, K.
(2003): In vitro activity of Brazilian medicinal plants,
naturally occurring naphthoquinones and their analogues,
against methicillin resistant Staphylococcus aureus. Int.
J. Antimicrob. Agents., 21, 279-284.
Maguire, H.; Pharoah, P.; Walsh, B. and Davison, C.
(2000): Hospital outbreak of Salmonella virchow possibly
associated with a food handler.J.hosp. infect.,44(4),2616.
Mahmoud, A.A.E. (1973): Changes in nutritive values of
gamma irradiated horticulture fruits during different
storage periods. M.Sc. Thesis, Fac. Agric., Ain Shams
Univ.
Mahrour, A.; Caillet, S.; Nketsia-Tabiri, J.and Lacroix, M.
(2003): Microbial and sensorial quality of marinated and
irradiated chicken. J. Food Prot., 66 (11), 21562159.
Mahrour, A.; Lacroix, M.; Nketsa-Tabiri, J.; Calderon, N.
and Gagnon, M. (1998): Antioxidant properties of
natural substances in irradiated fresh poultry.
Radiat.Physi Chem., 52, 77-80.
Malheiros, P.; Passos, C.; Casarin, L.; Serraglio, L. and
Tondo, E. C. (2010): Evaluation of growth and transfer
of Staphylococcus aureus from poultry
meat
to
surfaces of stainless steel and polyethylene and their
disinfection. Food Control., 21(3), 298-301.
192
Introduction
References
Markovic, K.; Hruskar, M. and Vahcic, N. (2006): Lycopene

content of tomato products and their contribution to the
lycopene intake of Croatians. Nutr. Res., 26, 556-560.
Martins, C.G.; Behrens, J.H.; Destro, M.T.; Franco,
B.D.G.M.; Vizeu, D. M.; Hutzler, B. and Landgraf, M.
(2004): Gamma radiation in the reduction of Salmonella
spp.Inoculated on minimally processed watercress
(Nasturium officinalis). Radiat.Phys.Chem., 71, 8993.
Martin-Salces, M.; de Paz, R.; Canales, M.A.; Mesejo,A.;
and
Hernandez-Navarro,F.(2008):Nutritional
recommend-dations in hematopoietic stem cell
transplantation. Nutrition., 24, 769775.
Marx, M.; Schieber, A. and Carle, R. (2000): Quantitative
determination of carotene stereoisomers in carrot juices
and vitamin supplemented (ATBC) drinks. Food Chem.,
70, 403408.
Mendoza, S.; Montemayor, L.; Boscan, L.A. and Barreiro,
J.A.(1982): Microflora in pasteurized fruit juices in
Venezuela. Arch. Latinoam. Nutr., 32 (3), 617629.
Mercier, J. and Lindow, S.E., (2000): Role of leaf surface
sugars in colonization of plants by bacterial epiphytes,
Appl, Environ. Microbiol., 66(1), 396-374.
Miean, K.H. and Mohamed, S. (2001): Flavonoid (myricetin,
quercetin, kaempferol, luteolin, and apigenin) content of
edible tropical plants. J. Agric. Food Chem., 49, 31063112.
Miguel, G.; Fontes, C.; Antunes, D.; Neves, A. and Martins,
D.(2004): Anthocyanin concentration of Assaria
pomegranate fruits during different cold storage
conditions. Journal of Biomedicine and Biotechnol., 338
342.
193
Introduction
References
Miller, W.R. and McDonald, R.E. (1996): Postharvest quality

of GA-treated Florida grapefruit after gamma irradiation
with TBZ and Storage, Postharvest Biol.Technol., 7, 253260.
Min, S.; Jin, Z.T.; Zhang, Q.H. (2003): Commercial scale
pulsed electric field processing of tomato juice. J. Agric.
Food Chem. 51, 3338-3344.
Mitchell, G.E.; Mclauchlan, R.L.; Isaacs, A.R.; Williams,
D.J. and Nottingham, S.M. (1992): Effect of low dose
irradiation on composition of tropical fruits and
vegetables. Journal of Food Composition and Analysis.,
5, 291-311.
Monk, J.D.; Beuchat, L.R. and Doyle, M.P. (1994):
Irradiation inactivation of foodborne microorganisms,
J.Food Prot., 58(2), 197-208.
Morehouse, K.M. (2002): Food irradiation US regulatory
considerations. Rad. Phys. Chem., 63, 281-284.
Mossel, D.A.A. and Degroot, A.P. (1965): The use of
pasteurizing doses of gamma radiation for the destruction
of Salmonella and other Enterobacteriaceae in some
foods of low water activity in: radiation preservation of
foods public. no.1273. NAS/NRS. Washington , D.C.P.
233-264.
Mossel, D.A.A. (1978): Streptococci in food. In : Streptococci
(Society for applied bacteriology, symposium series No.7)
(Skinner F.A. and Quesnel L.B. eds.), London : Academic
Press. P.392.
Moy, J.H. and Wong, L. (1996): Efficacy of gamma radiation
as a quarantine treatment of fruits, herbs and ornamentals
for Hawaii. Rad.Phys. Chem., 48, 373-374.
194
Introduction
References
Mukherjee, A.; Speh, D.; Jones, A.T.; Buesing, K.M. and

Diez-Gonzalez, F. (2006): Longitudinal microbiological
survey of fresh produce grown by farmers in the upper
Midwest. Journal of Food Protection 69, 19281936.
Narvaiz, P.; Lescano, G.; and Kairiyama, E. (1992):
Irradiation of almonds and cashew nuts, Lebensm. Wiss.
Technol., 25, 232-235.
Nguyen-the, C. and Carlin, F. (1994): The microbiology of
minimally processed fresh fruits and vegetables. Critical
Reviews in Food Sci and Nutr., 34, 371401.
Nguyen, M.L. and Schwartz, S.J. (1999): Lycopene: chemical
and biological properties. Food Technol., 53(2), 3845.
Niemira, B.A. and Deschnes, L. (2004): Ionizing radiation
processing of fruits and fruit products. In D.M. Barrett, L.
Somogyi, and H. Ramaswamy (Eds.), Processing fruits
(2nd ed.). Boca Raton, FL: CRC press.
Niemira, B.A.; Sommers, C.H. and Boyd, G. (2001):
Irradiation inactivation of four Salmonella species in
orange juices with varying turbidity. J. Food Prot., 64 (5),
614617.
Niven, C.F.Jr. (1985): Microbiological aspects of radiation
preservation of food. Ann. Rev. Microbiol., 12, 597-601.
Njobeh, P.B; Dutton, M.F.; Koch, S.H.; Chuturgoon, A.;
Stoev, S. and Seifert, K. (2009): Contamination with
storage fungi of human food from Cameroon, Int. J. Food
Microbiol., 135, 193198.
195
Introduction
References
Noci, F.; Riener, J.; Walkling-Ribeiro, M.; Cronin, D.A.;

Morgan, D.J. and Lyng, J.C. (2008). Ultraviolet
irradiation and pulsed electric fields (PEF) in a hurdle
strategy for the preservation of fresh apple juice, J. Food
Engine., 85, 141-984.
Nutraingredients USA. (2007): Global carotenoid market to hit
a new high.
http://www.nutraingredients.com/news/printNewsBis.asp
?id=78487
Olson, D.G. (1998): Irradiation of Food. Food Technol.,52, 5662.
Olsen, S.J.; MacKinon, L.C.; Goulding, J.S. and Slutsker,
L.(2000): Surveillance for foodborne disease outbreaks
United States,19931997. Morbidity and Mortality
Weekly Report 49, 151.
O'Neill, K.; Damoglou, A.P. and Patterson, M.F. (1991):
Sensitivity of some common grain fungi to irradiation on
grain and in phosphate-buffered saline. Letters in Applied
Microbiol., 12,180-183.
Orabi, I.O.A. (1981): Effect of gamma radiation and growth
regulators on chemical composition, yield and quality of
potatoes. M.Sc. Thesis, Fac. Agric., Ain Shams Univ.
Oussalah, M.; Caillet, S. and Lacroix, M. (2006): Mechanism
of action Spanish oregano, Chinese cinnamon, and savory
essential oils against cell membrane and walls of E. coli
O157:H7 and Listeria monocytogenes. J.of Food Prot.,
69, 10461055.
Paesmans, M. (2000): Risk factors assessment in febrile
neutropenia. Int. J. of Antimicrobial Agents, 16, 107-111.
196
Introduction
References
Palumbo, S.A.; Maximo, F.; Williams, A.C.; Buchanan, R.L.

and Thayer, D.W. (1985): Starch ampicillin agar for the
quantitative detection of Aeromonas hydrophila. Appl.
Environ. Microbiol., 50, 1027-1030.
Parish, M.E. and Higgins, D.P. (1989): Yeasts and molds
isolated from spoiling citrus products. J. Food Prot., 52,
261263.
Park, S.J.; Lee, J. I. and Park, J. (2002): Effects of a
combined process of high-pressure carbon dioxide and
high hydrostatic pressure on the quality of carrot juice. J.
Food Sci., 67(2), 18271834.
Patil, B.S.; Vanamala, J. and Hallman, G. (2004): Irradiation
and storage influence on bioactive components and
quality of early and late season Rio Red grapefruit
(Citrus Paradisi Macf.). Posthar. Bio.Technol., 34, 53-64.
Patterson, M.F. (1988): Sensitivity of bacteria to irradiation on
poultry meat under various atmosphere letters, Appl.
Microbiol., 7, 55.
Paull, R.E. (1999): Effect of temperature and relative humidity
on fresh commodity quality. Posthar.Bio.Technol., 15,
263-277.
Pickett, K.T.; Foley, D.; Caporaso, F.; Vasconcellos, A.;
Prakash, A. (2000): Effects of low dose irradiation on
the microbial counts , sensory attributes and ascorbic acid
content of unpasteurized orange juice, Abstract 86H-10,
Institute of Food Technologists Annual Meeting, 2000
http://ift.confex.com/ift/2000/techprogram/paper_3867.ht
m.
197
Introduction
References
Pizzo, P.A.; Purvis, D.S. and Waters, C. (1982): Microbiological evaluation of food items. For patients
undergoing
gastrointestinal
decontamination
and
protected isolation. J Am. Diet. Assoc., 81, 272279.
Polidori, M.C. (2003): Antioxidant micronutrients in the
prevention of agerelated diseases. J. Postgrad. Med., 49,
229235.
Ponniah, J.; Robin, P.; Paie, M.S.; Radu, S.; Ghazali, F.M.;
Kqueen, C.Y.; Nishibuchi, M.; Nakaguchi, Y. and
Malakar, P.K. (2010): Listeria monocytogenes in raw
salad vegetables sold at retail level in Malaysia , Food
Control., 21, 774778.
Prakash, A.; Inthajak, P.; Huibregtse, H.; Caporaso, F. and
Foley, D. (2000): Effects of low-dose gamma irradiation
and conventional treatments on shelf life and quality
characteristics of diced celery. J Food Sci., 65, 1070
1075.
Prakash, A.; Manley, J.; Decosta, S.; Caporaso, F.; Foley, D.
(2002): The effects of gamma irradiation on the
microbiological, physical and sensory qualities of diced
tomatoes. Radiat.Phys.Chem., 63, 387-390.
Pryke, D.C. and Taylor, R.R. (1995): The use of irradiated
food for immunosuppressed patients in the United
Kingdom. J.Human Nutr.Diet., 8, 411416.
Przybylska, A. (2001): Outbreak of foodborne and waterborne
infections and intoxications in hospitals and sanatoria in
Poland in 19851999. Przeglad epidemiologiczny, 55(1
2), 21729 (abstract) [In Polish].
Rao, A.V. and Agarwal, S. (1999): Role of lycopene as
antioxidant carotenoid in the prevention of chronic
diseases: A Rev.Nutr.Res.,19, 305-323.
198
Introduction
References
Reglier-Poupet, H.; Parain, C.; Beauvais, R.; Descamps, P.;

Gillet, H.; Le Peron, J.; Berche, P.; Ferroni, A.(2005):
Evaluation of the quality of hospital food from the kitchen
to the patient. Journal of hospital infection., 59(2),1317.
Rendueles, E.; Omer, M.K.; Alvseike, O.; Alonso-calleja, C.;
Capita, R. and Prieto, M.J. (2011): Microbiological
food safety assessment of high hydrostatic pressure
processing: A review, LWT - Food Sci.Technol.,44, 12511260.
Rentzsch, M.; Schwarz, M. and Winterhalter, P. (2007):
Pyranoanthocyanins an overview on structures,
occurrence, and pathways of formation. Trends in Food
Sci. and Technol., 18(10), 526-534.
Risi, G.F. and Tomascak, V. (1998): Prevention of infection in
the immunocompromised host, Am J Infect Control., 26,
594-606.
Rodriguez, M.; Valero, A.; Carrasco, E.; Prez-Rodrguez,
F.; Posada, G.D. and Zurera, G.(2011) : Hygienic
conditions and microbiological status of chilled ReadyTo-Eat products served in Southern Spanish hospitals,
Food Control., 22, 874-882.
Roilides, E.; Lamaignere, C.G.; and Farmaki, E.(2002):
Cytokines in immune deficient patients with invasive
fungal infections: an emerging therapy. Int.J Infect.Dis.,
6, 154-163.
Roldan Gutierrez, J.M. and Luque de castro, M.D. (2007):
Lycopene: the need for better methods for
characterization and determination, Trends in Analytical
Chemistry. 26(2), 154-162.
199
Introduction
References
Rowley, D.B.; Sullivan, R. and Josephson, E. (1978):

Indicator of viruses in foods preserved by ionizing
radiation. Indicator of viruses in water of food, edited by
Gerald Berg. Ann.Arbor Science . publishers INC
Copenhagen.
Ryu, J.H. and Beuchat, L.R. (1998): Influence of acid
tolerance responses on survival, growth and cross
protection of Escherichia coli O157:H7 in acidified media
and fruit juices. Int. J.Food Microbiol., 45,185-193.
Safadi, A. and Soubani, A.O. (2009): Diagnostic approach of
pulmonary disease in the HIV negative immunocompromised host. Eur.J.Int.Medicine., 20, 268279.
Saldana, G.; Stephens, T.S. and Lime, B.J. (1976): Carrot
beverages. J. Food Sci., 41 (5), 12431244.
Saltveit, M.E. (1997): Physical and physiological changes in
minimally processed fruits and vegetables. In: TomasBarberan, F.A. (Ed.), Phytochemistry of Fruit and
Vegetables. Oxford University Press, Oxford, UK, pp.
205 220.
Sandmann, G. (1994): Carotenoid biosynthesis in microorganisms and plants. Eur. J. Biochem. 223, 7-24.
SantAna, H.M.P.; Stringheta, P.C.; Brandao, S.C.C. and
De-Azeredo, R.M.C. (1998): Carotenoid retention and
vitamin A value in carrot ( Daucus carota L. ) prepared
by food service. Food chem., 61, 145-151.
SAS (1997): SAS / STAT user, version 6.12 TS020 by SAS
Institute Inc., Cary, Licensed to Penn State University,
Site 0016309001.
200
Introduction
References
Sato, T., and Miyata, G. (2000): The nutraceutical benefit, part

iv: garlic. Nutrition., 16, 787788.
Saura-Calixto, F. and Goi, I. (2009): Definition of the
Mediterranean diet based on bioactive compounds. Crit.
Rev.Food Sci.Nutr., 49, 145152.
Schlech, W.F.; Lavigne, P.M.; Bortolussi, R.A.; Allen, A.C.;
Haldane, E.V.; Wort, A.J. and Hightower, A.W.
(1983): Epidemic listeriosis-evidence for transmission by
food. New England Journal of Medicine., 308, 203206.
Schwartz, S.A. and Perry, S. (1966): Patient protection in
cancer chemotherapy. JAMA , 197, 623627.
Sharon Raber, O. and Kahn, V. (1983): Avocado
Monocarp;
browning
potential
carotenoid
content,polyphenol oxidase, catalase and peroxidase
activity comparsion between six avocado cultivars. J.
Food Sci., 48, 1874-1875.
Shelef, L.A. (1983): Antimicrobial effects of spices. J. Food
Safety., 6, 29-44.
Shengfu, Y.; Yuanhu, Z.; Bingsong, C. and Suqin, Z. (1993):
Effects of cobalt-60 -ray irradiation on fresh keeping and
storage of kiwifruits. Radiat.Phys.Chem. 42(1-3), 339341.
Shi, J. and Le Maguer, M.(2000): Lycopene in tomatoes:
chemical and physical properties affected by food
processing. Crit.Rev.Food Sci.Nutr., 40(1),142.
Shi, J.; Le Maguer, M.; Kakuda, Y.; Liptay, A. and
Niekamp , F. ( 1999): Lycopene degradation and
isomerisation in tomato dehydration . Food Res.Int., 32
(1), 15-21.
201
Introduction
References
Shibamoto, T.; Bjeldanes, L.F. and Taylor, S. (1993):

Introduction to Food Toxicology. Academic Press.
Singh, H. (1990): Nutritional aspects of irradiated Mangoes.
Atomic Energy of Canada Ltd. Pinawa, M.B. (Canada).
Whites hell Nuclear Research Establishments, Jun, 36, p.
Sizer, C.E. and Balasubramaniam, V.M. (1999): New
intervention processes for minimally processed juices,
Food Technol., 53(10), 64-67.
Smid, E.J. and Gorris L.G.M. (1999): Natural antimicrobials
for food preservation. In: "Handbook of food
preservation" (Rhaman M.S.), Marcel Dekker, New York.
Smith-Palmer, A.; Stewart, J. and Fyfe, L. (1998): Antimicrobial properties of plant essential oils and essences
against five important food-borne pathogens. Lett. Appl.
Microbiol., 26, 118-122.
Soliman, K.M. and Badeaa, R. I. (2002): Effect of oil
extracted from some medicinal plants on different
mycotoxigenic fungi. Food and Chemical Toxicology, 40,
1669-1675.
Somerville, E. T.(1986): Special diets for neutropenic patients:
do they make a difference? Semin.Oncol.Nurs., 2(1), 5558.
Sommer, N.F. (1973): The effect of ionizing radiation on fungi.
Manual of radiation sterilization of medical and biological
materials. IAEA, Vienna , 73.
202
Introduction
References
Somogyi, L.P. and Romani, R.J. (1964): Irradiation induced

textural change in fruits and its relation to pectin
metabolism. J. Food. Sci., 29, 366-371.
Song. H.P.; Byun. M.W.; Jo.C; Lee.C.H; Kim. K.S and
Kim. D. H. (2007): Effects of gamma irradiation on the
microbiological, nutritional, and sensory properties of
fresh vegetable juice, Food Control., 18, 510.
Spinks, J.W.T. and Woods, R.J. (1990): Organic compounds.
In: An Introduction to Radiation Chemistry, Third
Edition. A Wiley-Interscience Publication, New York, pp.
392396 (Chapter 9).
Spoto, M.H.F.; Domarco, R.E.; Walder, J.M.M.; Scarminio,
I.S. and Bruns, R.E. (1997): Sensory evaluation of
orange juice concentrate as affected by irradiation and
storage, J.Food Process, Preserv., 21(3),179-191
Stahl, W. and Sies, H. (1996): Lycopene : a biologically
important carotenoid for humans? Arch. Biochem.
Biophys. 336, 1-9.
Stevenson, M.H. (1994): Progress in the identification of
irradiated foods. Food Technol., 48, 141-144.
Sukhwant, M.K.; Harvinder, K. and Tejinder, G. (1992):
Effect of cooking on fibre content of vegetables. J. Food
Sci.Technol, 29, 185186.
203
Introduction
References
Swailam, H.M.H. (1998): Preservation of Mango and Mango

pulp by radiation, P.H.D. Thesis, Fac. Agric. Suez Canal
Univ. Cairo, Egypt.
Tannenbaum, S.R.; Young, V.R. and Acher, M.C. (1985):
Vitamins and minerals. In: Fennema, O.R.(Ed.), Food
Chem., Marcel Dekker, Inc., New york, USA, pp. 488493.
Talcott, S.T.; Brenes, C.H., Pires, D.M. and Del PozoInsfran, D. (2003): Phytochemical stability and color
retention of copigmented and processed muscadine grape
juice. J. Agric. Food Chem., 51, 957963.
Tauxe, R.; Kruse, H.; Hedberg, C.; Potter, M.; Madden,
J.and Wachsmuth, K. (1997): Microbial hazards and
emerging issues associated with produce. A preliminary
report to the national advisory committee on microbiological criteria for foods. J. Food Protect., 60, 1400
1408.
Tehranifar, A.; Zarei, M.; Nemati, Z.; Esfandiyari, B. and
Vazifeshenas, M.R. (2010): Investigation of physiochemical properties and antioxidant activity of twenty
Iranian pomegranate (Puncia granatum L .) Cultivars.
Scientia Horticulturae., 126,180-185.
Teller, K.H. and Parish, M.H. (1992): Microbially produced
offflavors in orange juice. Proc.Fla.State Hortic. Soc.,
105, 144146.
204
Introduction
References
Thomas, P. (1986). Radiation preservation of foods of plant

origin. Part V. Temperate fruits: pome fruits, stone fruits,
and berries. In: CRC Critical Reviews in Food Sci. and
Nutr., Vol. 24(4). CRC Press, Boca Raton, FL, pp. 357
400.
Thornley, M.J. (1963): Radiation resistance among bacteria.
J.Appl. Bacteriol., 26, 334-345.
Tiwari, B.K.; O'Donnell, C.P. and Cullen, P.J. (2009): Effect
of non thermal processing technologies on the
anthocyanin content of fruit juices. Trends in Food Sci.
and Technol., 20(3-4), 137-145.
Tiziani, S. and Vodovotz, Y. (2005): Rheological effects of soy
protein addition to tomato juice. Food Hydrocoll., 19, 45
52.
Todd, J.; Schmidt, M.; Christian, J. and Williams, R.
(1999): The low-bacteria diet for immunocompromised
patients. Reasonable prudence or clinical superstition?
Cancer Pract., 7, 205-7.
Todoriki, S.; Bari, L.; Kitta, K.; Ohba, M.; Ito, Y.;
Tsujimoto, Y.; Kanamori, N .; Yano, E.; Moriyama,
T.; Kawamura, Y.; and Kawamoto, S. (2009): Effect of
gamma-irradiation on the survival of Listeria
monocytogenes and allergenicity of cherry tomatoes.
Radiat.Physi.Chem., 78, 619621.
205
Introduction
References
Torskangerpoll, K. and Andersen, Q.M. (2005): Colour

stability of anthocyanins in aqueous solutions at various
pH values. Food Chem., 89, 427440.
Tournas, V.H.; Heeres, J. and Burgess, L. (2006): Moulds
and yeasts in fruit salads and fruit juices. Food
Microbiol., 23, 684688.
Tuormaa, T. (1994): The Adverse Effects of Food Additives on
Health: A Review of the Literature with Special Emphasis
on Childhood Hyperactivity. Journal of Orthomolecular
Medicine. 9(4), 225-243.
Urbain, W.M. ( 1986 ): Biological effects of ionizing radiation.
In : Food Irradiation. Food Science and Technology
series. Academic press, London, pp. 83-117.
US Food and Drug Administration (USFDA), 2001./http://
www.cfsan.fda.gov/_comm/ift3-4a.htmlS(accessed
21.09.05).
Vachon, C.; DAprano, G.; Letendre, M. and Lacroix, M.
(2002): Effect of edible coating process and irradiation
treatment of strawberry Fragaria spp. On storage keeping
quality. J. Food. Sci., 68, 608612.
Valero, M. and Salmeron, M.C. (2003) Antibacterial activity
of 11 essential oils against Bacillus cereus in tyndallized
carrot broth, Int.J. Food Microbiol., 85, 7381.
206
Introduction
References
Van Duynhoven, Y.T.; de Jager, C.M.; Kortbeek, L.M.;

Vennema, H.; Koopmans, M.P.; van Leusden, F.; Van
der Poel, W.H. and Van der Broek (2005): A one-year
intensified study of outbreaks of gastroenteritis in The
Netherlands. Epidemiol Infect.133, 921.
Van tiel, F.H.; Harbers, M.M.; Terporten, P.H.W.; Van
boxtel, R.T.C.; Kessels, A.G.; Voss, G.B.W.E. and
Schouten, H.C. (2007): Normal hospital and low
bacterial diet in patients with cytopenia after intensive
chemo-therapy for haematological malignancy: a study of
safety. Annals of Oncol. 18, 1080-1084
Verhoeff-Bakkenes, L.; Jansen, H.A.P.M.; In 't Veld, P.H.;
Beumer, R.R.; Zwietering, M.H. and van Leusden,
F.M. (2011): Consumption of raw vegetables and fruits:
A risk factor for Campylobacter infections, Int. J. Food
Microbiol., 144, 406412.
Vlad, E.; Zarnescu, A.; Arizan, D.; Marseu, P. and
Macedon, T. (1971): Effect of heat treatment plus
gamma irradiation and gamma irradiation in the cold on
survival of Saccharomyces cerevisiae cells. Industeria
Alimentara. 22,(5), 261-264.Quote form FSTA, 4(8),
1972
207
Introduction
References
Voravuthikunchai, S.; Lortheeranuwat, A.; Jeeju, W.;

Sririrak, T.; Phongpaichit, S. and Supawita, T. (2004):
Effective medicinal plants against enterohaemorrhagic
E.coli O157:H7. Journal of Ethnopharmaacology .,94,
49-54.
Voutilainen, S.; Nurmi, T.; Mursu, J. and Rissanen, T.H.
(2006) : Carotenoids and cardiovascular health. Am. J.
Clin. Nutr., 83, 12651271.
Wade, W.N.; Vasdinnyei, R.; Deak, T.; Beuchat, L.R. (2003).
Proteolytic yeasts isolated from raw, ripe tomatoes and
metabiotic association of Geotrichum candidum with
Salmonella, Int. J.Food Microbiol., 86,101-111.
Waheed, S.; Siddique, N.; Rahman, A.; Zaidi, J.H. and
Ahmad, S. (2004): INAA for dietary assessment of
essential and other trace elements in 14 fruits harvested
and consumed in Pakistan. J of Radioanalytical and
Nuclear Chem., 260, 523531.
Wang, H.; Cao, G. and Prior, R.L.( 1996): Total antioxidant
capacity of fruits. J. Agric.Food Chem. 44,701705.
Wang, H.; Hu, X.; Chem, F.; Wu, J.; Zhang, Z.; Liao, X.;
Wang, Z.; (2006): Kinetic analysis of non-enzymatic
browning in carrot juice concentrate during storage. Eur.
Food Res.Technol., 223, 282289.
208
Introduction
References
Wang, S.; Melnyk, J.P.; Tsao, R.; Marcone, M.F. (2011):

How natural dietary antioxidants in fruits, vegetables and
legumes promote vascular health. Food Res. Inter., 44,
14-22.
Wani, A.M.; Hussain, P.R.; Meena, R.S. and Dar, M.A.
(2008): Effect of gamma-irradiation and refrigerated
storage on the improvement of quality and shelf life of
pear (Pyrus communis L., Cv. Bartlett/William). Radiat.
Phys. Chem., 77, 983 989.
Warnasuriya, D.; Liyanage, A.W.; Weerawansa, G.G.;
Athauda, P.K. and Jayatissa, P.M. (1985): Isolation
and characterization of yeasts of some fruits and fruit
products of Srilanka. J.Natn.Sci.Coun.Srilanka., 13(1).
WCRF, World Cancer Research Fund and American
Institute for Cancer Research (2007): Food, nutrition,
physical activity and the prevention of cancer: A global
perspective. Washington, DC, USA: American Institute
for Cancer Research.
Wells, J.M. and Butterfield, J.E. (1997): Salmonella
contamination associated with bacterial soft rot of fresh
fruits and vegetables in the market place. Plant Dis., 81,
867-872.
Wen, H.; Chung, H.; Chuo, F.; Lin, I. and Hsieh, P. ( 2006 ):
Effectof gamma irradiation on microbial decontamination
and chemical and sensory characteristic of Lycium fruit,
Rad. Phys. Chem., 75, 596-603.
209
Introduction
References
Wendakoon, C.N. and Sakaguchi, M. (1995): Inhibition of

amino acid decarboxylase activity of Enterobacter
aerogenes by active components in spices. J.Food
Protect., 58, 280283.
WHO (1988): Food Irradiation. A technical for preservation
and improving the safety of food. WHO, Geneva
WHO (1993): Guidelines for drinking-water quality. Vol
Recommendations, 2nd Edn. Geneva, World Health
Organization.
WHO (1997): Consultations and workshops: Prevention and
control of Enterohaemorrhagic Escherichia coli ( EHEC )
infection. Report of a WHO consultation, April 28-May 1,
1997. Food Safety Unit, Programme of Food safety and
Food Aid, Geneva.
WHO (1999): High-Dose Irradiation: Wholesomeness of food
irradiated with dose above 10 kGy: report of a Joint FAO/
IAEA/WHO Study Group. In: World Health Organization
Technical Report Series 890. WHO, Geneva, Switzerland,
7pp.
WHO (2000): Foodborne Disease: A focus for Health
Education.
Williams, C. (1995): Healty eating: clarifying advice about fruit
and vegetables. Brit. Med. J., 310, 1453-1455.
Wilson, B.J. (2002): Dietary recommendations for neutropenic
patients. Sem.Oncol.Nur.,18 (1), 4449.
Wimsen, P.K.; Spada, D.S. and Salavador, M. (2005):
Antioxidant activity of the flavonoid hesperidin in
chemical and biological systems. J. Agric. Food Chem.,
53, 4757-4761.
210
Introduction
References
Wood, O.B. and Bruhn, C.H., (1999): Food irradiation

position of the ADA (AmericanDieteticAssociation)
,http://www.eatright.org/airradi.html.
Wood, O.B. and Bruhn, C.M. (2000): Position of the
American dietetic association: food irradiation. J. Am.
Diet Assoc., 100, 246-253.
Wu, V.C.H.; Fung, D.Y.C. and Kang, D.H. (2001):
Evaluation of thin agar layer method for recovery of coldinjured foodborne pathogens. J. Rapid Meth. Auto.
Microbiol. 9, 1125.
Yahia, E.M. (2010): The contribution of fruits and vegetable
consumption to human health. In L. A. De la Rosa, E.
lvarez-Parilla and G. A. Gonzlez-Aguilar (Eds.), Fruit
and vegetable phytochemicals (pp. 351). Iowa: WileyBlackwell.
Yoon, K.Y.; Woodams, E.E. and Hang, Y.D. (2004):
Probiotication of tomato Juice by Lactic Acid Bacteria. J.
Microbiol., 4, 315-318.
Young, E. (1997): Prevalence of intolerance to food additives.
Environ.Toxicol.Pharmacol., 4,111114 .
Young, L.S. and Rubin, R.H. (1994): Introduction. In: Rubin
RH, Young LS, editors. Clinical approach to infection in
the compromised host. 3rd ed. New York: Plenum; 1994.
p. 1-5.
Youssef, B. M.; Asker, A. A.; El-samahy, S.K. and Swailam,
H.M. (2002): Combined effect of steaming and gamma
irradiation on the quality of mango pulp stored at
refrigerated temp. Food Res. Int., 35, 1-13.
211
Introduction
References
Zaika, L.L. (1988): Spices and herbs their antimicrobial

activity and its determination. J. Food Saf., 9, 97-118.
Zegota, H. (1999): The effect of gamma irradiation on citrus
pectin in N2Oand N2O/O2 saturated aqueous solutions.
Food Hydrocolloid. 13, 51-58.
Zhang, D. and Hamauzu, Y. (2004): Phenolics, ascorbic acid,
carotenoids and antioxidant activity of brocolli and their
changes during conventional and microwave cooking.
Food Chem., 88, 503509.
Zhang, L-X.; Conney, R.V. and Bertram, J.S. (1991):
Carotenoids enhance gap junctional communication and
inhibit lipid peroxidation in C3H/1077/2 cells:
relationship to their cancer chemopreventative action.
Carcinogenesis, 12, 2109-2114.
Zhang, L.; Lu, Zhaoxin, L. and Wang, H. (2006): Effect of
gamma irradiation on microbial growth and sensory
quality of fresh-cut lettuce. Int. J. Food microbiol.,106,
348-351.
212

) (
)(
:
:

:
-. / . ......................................
.
-. /. ........................................

.
- / . .........................................
-
- .

:
:

:
-. / . :
.
-. /. :
.
- / . : -
-
.
:
-./. :
.
-. /. :
.
-. / . :
.
-. /. :
.
/ / :
:

:

/ /
/

/ /
:
:
:
:
:
:
:
.

.
. /. :

.
. /. :
.

.
/. :

.
.
:
:
:
:
:
:

.

.
.
) - (
) (

.

.
) (
.
/

.

-

.
.
)
(
.
) - -
(

.
--
:
:
-
.
- )
( .
-
)
.( D10value
-

.
-
.
-
.
:
- )
(
) Candida tropicalis (
) Debaryomyces hansenii (
) Saccharomyces cerevisiae ( Trichosporon
)pullulans ( ) Rhototorulaglutinis
( .
--
-
Candida tropicalis, Rhodotorula glutinis, Saccharomyces
cerevisiae , Trichosporon pullulans and Debaryomyces
hansenii

.
-
/
/ .
/
/

/
/
.
-

) (
) (
. %
- .

0.5

.
) (

) ( ,
--

.
- .

) .(
-
/

) .(
--

Development of Safe Juices For Immunocompromised Patients by Irradiation Alone or in Combination With Other Technologies

Transféré par

Informations du document

Titre original

Copyright

Formats disponibles

Partager ce document

Partager ou intégrer le document

Options de partage

Avez-vous trouvé ce document utile ?

Ce contenu est-il inapproprié ?

Droits d'auteur :

Formats disponibles

Development of Safe Juices For Immunocompromised Patients by Irradiation Alone or in Combination With Other Technologies

Transféré par

Droits d'auteur :

Formats disponibles

FacultyofScience

Development of safe juices for

Reham Mohamed Mohamed Abd El-kader

Development of safe juices for

Reham Mohamed Mohamed Abd El-Kader

Prof. Khayria Abd El-Ghany Youssef

Prof. Ali Ahmed Ibrahim Hammad

Dr. Hanan Hassan Abd El-Khalek

Name: Reham Mohamed Mohamed Abd El-Kader

1-Prof. Khayria Abd El-Ghany Youssef

2-Prof. Ali Ahmed Ibrahim Hammad..

3- Dr. Hanan Hassan Abd El-Khalek.

Development of safe juices for

Reham Mohamed Mohamed Abd El-Kader

1-Prof. Mohie Eddin Zohier El-Fouly

2-Prof. Mohamed Ghareeb Ibrahim Emaish

3-Prof. Khayria Abd El-Ghany Youssef

4-Prof. Ali Ahmed Ibrahim Hammad

My deepest graditude to all staff of Microbiology department,

Name : Reham Mohamed Mohamed Abd El-Kader

2. Immunocompromised patients problems

3. Immunocompromised patients recommended diets

4.1. Pomegranate juice

4.2. Tomato juice

4.3. Carrot juice

4.7. Ascorbic acid

5. Microbial contamination of food in hospitals

6. Microbial contamination of fresh fruits and vegetables

7. Microbial contamination of fresh fruits and vegetables juices

8. Methods used to reduce the hazards of foodborne infection for

8.1. Thermal treatments

8.3. Chemical additives

8.4. High hydrostatic pressure processing ( HHP )

8.5. Ionizing radiation

9. Treatment of food by irradiation

9.1. Radiation sources used for food irradiation

9.2. Wholesomeness and safety aspects

9.3. International approval

9.4. Technological benefits and advantages

10. Irradiation effects

10.1. Relative radiation resistance of microorganisms

10.2. Effect of irradiation on microbial load of fresh

10.3. Effect of irradiation on foodborne pathogenic

10.4. Effect of irradiation on microbial quality of fruits

10.5. Effect of irradiation of the main chemical

10.5.4. Ascorbic acid

10.6. Effect of irradiation on sensory properties

10.7. Viscosity effect

11. Preservation of juices by irradiation and other

11.2. Irradiation in combination with natural additives

Materials and Methods

1.1. Preparation of fresh squeezed pomegranate juice

1.2. Preparation of fresh squeezed tomato juice

1.3. Preparation of fresh squeezed carrot juice

4.1. Total aerobic bacterial counts ( TBC)

4.2. Lactic acid bacteria ( LAB)

4.3. Total molds and yeasts

4.4. Total coliform

4.5. Escherichia coli

4.6. Enterococcus faecalis

4.7. Staphylococcus aureus

4.8. Aeromonas hydrophila

5. Isolation and purification of yeasts