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SESIPENGAJIAN: 'd-0 0t5/0b hJ()3a Ct .J.ODe)oCJ

(HURUF BESAR)

n.~t·H?;aku membenarkan tesis (LPS/ Sarjana/ Doh1or Falsafah) ini di simpan di Perpustakaan Universiti Malaysia Sabah ~ttgan syarat-syarat kegunaan seperti berikut:

I. Tesis adalah halanilik Universiti Malaysia Sabah. 'J PeI}>ustakaan Universiti Malaysia Sabah dibenarkan membuat salinan untuk tujuan pengajian sahaja. 3. PeI}>ustakaan dibenarkan membuat salin an tesis ini sebagai bahan pertukaran antara ins[i tusi pengajian tinggi.

4. ** Si1a tandakan ( / )

SULIT

TERHAD

/' J ,TIDAK TERHAD

~ (TAND~PENULIS) ~qtnflt Tetap: NO-S) Loru~q thvCU) If /

~()cJt:'I ~yy)p M*~.J-..o!/~ ___ _

~ 6 8- 000 4-Mpotn ff I IS 01 Me C) f

't--AT AN: * Potong yang tidak berkenaan.

(Mengandungi maldumat yang berdarjah keselamatan alau kepentingan Malaysia seperti yang tem1aktub di dalam AKT A RlillSIA RASMI 1972)

(Mengandungi maklumat TERHAD yang telah ditenhlkakan oleh organisasilbadan di mana peny lidikan dijalankan)

Disahkan oleh

(TA iDATANGAN PU TAKAWAN)

'Prof · f'nad.!:JOi Dr . C""'!:1<L Fool: Ye.~ Nama Penyeha

Tarikh: I~ c) c to b Q..r ';>"009 -- - ..

* Jika tesis ini SULIT atau TERHAD, si1a lampiran surat daripada pihak berkuasa/organsasi berkenaan dengan menyatakan seka!i sebab dan tempoh tesis ini perlu dike laskan sebagai SULIT

danTE~AD. .. . ./ * Tesls dlmaksudkan sebagal teslS bagl ~azah Doktor Falsafah dan Sarjana secara penyeJiaikan, atau

diserta?1 bagi pengajian secara kerja kur sus dan penyelidikan, atau Laporan Proj ek arjana Muda (LPSM).

" / -

GROWTH AND SURVIVAL OF PROBIOTICS (Lactobacillus caseiand Lactobacillus acidophilus) IMMOBILIZED ON

DEHYDRATED FRUIT CUBES

TANYEE MUN

PERPlSTAUAN ,Fasm MALAYSIA SARAH

THIS THESIS IS SUBMITTED TO FULFILL PARTIAL REQUIREMENTS OF THE DEGREE OF BACHELOR

OF FOOD SCIENCE WITH HONOURS (FOOD SCIENCE AND NUTRITION)

SCHOOL OF FOOD SCIENCE AND NUTRITION UNIVERSITI MALAYSIA SABAH

2009

DECLARATION

I hereby declare that the materials in this thesis are the result of my own research accept as stated in references.

17 April 2009

11

APPROVAL

CERTIFIED BY

SIGNAlURE

1. SUPERVISOR (PROF. MADYA DR. CHYE FOOK YEE)

2. EXAMINER-l (DR. PATRICIA MATANJUN)

3. EXAMINER-2 (Ms. HO AI UNG)

4. DEAN (PROF. MADYA DR. MOHO. ISMAIL ABDULLAH)

3Jld-.

iii

ACKNOWLEDGEMENT

First of all, I want to thanks my sincere thanks to my supervisor, Prof. Madya Dr. Chye Fook Vee for his guidance throughout the whole research. Not only had that he graciously supplied some research materials and kind enough to comment helpfully on my research and but also stated out the mistakes that had been made for a better improvement.

I would like to express gratitude to my parents in terms of financial and moral support. Also to all my beloved friends who had help me in this entire research and giving helping hand throughout the whole process of completing this research. Their countless support and help will be much appreciated.

Last but not least, to many individuals who have in their own special ways contributed to this research.

iv

ABSTRACT

In this study, the effect of different methods of fermentation, fruits, sizes of fruit cubes, dehydration methods and storage temperatures on the growth and survival of L. acidophilus and L. casei immobilized on fruit cubes were evaluated. Results indicated that L. acidophilus had significant (p<0.05) higher peak growth in papaya, banana and pineapple fruit juices than L. case~ with the viable cell numbers of 8.35 log cfu g-t, 8.34 log cfu g-1 and 8.21 log cfu g-1 respectively. Immobilization of culture on fruit cubes immersed into its respective fruit juices inoculated with culture was studied, with immersion of fruit cubes prior to fermentation and after fermentation. Proven that there was a significant (p<0.05) higher growth of immobilized probiotic cells which went through the immersion of fruit cubes along fermentation. The immobilization study also showed that, fruit cubes sized 1.0 cm3 had better growth significantly (p<0.05) of L. acidophilus or L. casei rather than on 1.5 cm3 fruit cubes. The pH value of immobilized fruits cubes decreased Significantly at the Initial point to the peak point of fermentation but only decrease slightly after the peak hour. The major changes of pH value may due to the accumulation of lactic acid produced from the metabolic pathway of growth. Freeze drying seems to be more effective in preserving the high viability of immobilized cultures and had viable cell number in the same logarithmic cycle even after the freeze drying process. Lastly as a conclUSion, the storage at -20°C for freeze dried 1.0 cm3 immobilized fruits cubes which went through the fermentation method of immersion along fermentation had the most favorable survival rate with the mean value of 7.52 log cfu g-l, 7.98 log cfu g-1 and 6.39 log cfu g-1 for papaya fruits, banana and pineapple fruits respectively.

v

ABSTRAK

PERnJHBUHAN DAN KEUPAYAAN UNnJK HIDUP PROBIOTIK YANG DINYAHHOBIUSASIKAN PADA KIUB BUAH-BUAHAN KERING

Dalam penyelidikan ini. kesan penggunaan cara fermentasi yang berlalnan, buah­buahan, saiz kiub buah-buahan, cara pengeringan and suhu penyimpanan terhadap pertumbuhan dan keupayaan untuk hidup L acidophilus and L casei yang dinyahmobilisasikan pada permukaan kiub buah-buahan dikaji. Keputusan menunjukkan L. acidophilus mempunyai perbezaan signifikan (p<0.05) pertumbuhan maximum yang leblh tinggl dalam betik, pisang dan nanas dibandingkan dengan L case~ dengan bacaan min 8.35 log cfu g-1, 8.34 log cfu g-1 and 8.21 log cfu g-1 masing-masing. Penyahmobillsasi kultur pada permukaan kiub buah-buahan dengan cara fermentas~ penambahan kiub buah sekall dengan fermentasl kultur dan penambahan pada tahap maximum fermentasi dikaji. Terbuktl terdapat pertumbuhan yang lebih tinggi secara signitikan (p<O.05) dengan cara fermentasi penambahan klub buah sekall dengan fermentas; kultur. Kiub bersaiz 1.0 orr mempunyai pertumbuhan L acidophilus or L casei yang lebih tinggi berbanding dengan kuib bersaiz 1.5 cw. Bacaan pH menurun apabila wujudnya pertumbuhan drastik kultur yang dinyahmobilisasikan pada permukaan kiub. Ini berlaku kerana pada pertumbuhan yang tingg~ penghasilan asid laktik meningkat aklbat proses metabollk kultur. Pengerlngan sejukbeku leblh efektlf dalam proses pengeringan dengan bacaan min yang maslh berada dalam logaritma yang sama selepas pengeringan. Kesimpulannya, kultur yang dinyahmobllisasikan dengan kaedah penambahan kiub sekali dengan fermentasi pada permukaan kiub bersaiz 1.0 CM yang melalui pegeringan sejuk beku disimpan pada suhu -2tfC mempunyai pengiraan koloni hldup yang paling tinggi dalam ketiga-tiga buah dengan bacaan min 7.521ogcfu g-1, 7.98 log cfu g-1 and 6.39 log cfu g-1 bagl buah betik, pisang dan nanas masing-",asing.

vi

TABLE OF CONTENT

TITLE

DECLARATION

APPROVAL

ACKNOWLEDGEMENT

ABSTRACT

ABSTRAK

TABLE OF CONTENT

LIST OF TABLE

LIST OF FIGURE

LIST OF SYMBOLS

LIST OF APPENDIX

CHAPTER 1 INTRODUCTION

CHAPTER 2 LITERATURE REVIEW 2.1 Roles of Functional Foods to Public Health

2.1.1 The Development of Functional Foods 2.1.2 Application of Bacteria in Functional Foods 2.1.3 Probiotics in Foods

a. Application of Probiotics in Healthcare Product b. Application of Probiotics in Dairy Products c. Application of Probiotics in Non-dairy Products

2.2 Benefits of Probiotics 2.1.1 Probiotics Benefits Health by Affecting Immune System 2.2.2 Probiotics Benefits Health by Strengthening the Mucosal 2.2.3 Probiotics Benefits Health by Strengthening Competitive

Exclusion 2.2.4 Probiotics Benefits Health by Suppressing Intestinal

Inflammation 2.3 Selection of Probiotics

2.3.1 Safety Aspects in Selection 2.3.2 Functionality Aspects in Selection

a. Adhesion Properties of Probiotics b. Immunomodulatory Properties of Probiotics c. Antagonistic Properties d. Antimutagenic and Anticarcinogenic Properties

2.3.3 Technological Aspects in Probiotics Selection 2.4 Challenges in Producing Foods with Probiotics

vii

Pages

ii

iii

iv

v

vi

vii

x

xi

Xiii

xiv

1

5 5 6 10 12 13 14 14 15 17

Barrier 18

19

19

20 21 22 23 24 24 25 26 27

2.4.1 Survival of Probiotics 2.4.2 Resistance of Probiotics towards Dehydration Process

a. Freeze Drying b. Oven drying

2.4.3 Storage of Probiotics 2.5 Immobilization of Probiotics into Fruit Matrix

2.5.1 Fruits 2.5.2 Banana 2.5.3 Papaya 2.5.4 Pineapple

CHAPTER 3 MATERIALS AND METHODS 3.1 Materials 3.2 Preparation of Cultures 3.3 Preparation of Fruit Medium 3.4 Preliminary Fermentation Trial of Strains in Fruit Juices Medium 3.5 Preliminary Immobilization Trial of Strains on Fruit Cubes 3.6 Immobilization of Strains on Fruit Cubes 3.7 Dehydration

3.7.1 Oven Dehydration 3.7.2 Freeze Drying

3.8 Rehydration of fruit cubes 3.9 Viability Test

3.9.1 Viability Test on Inoculated Medium 3.9.2 Viability Test on Immobilized Fruit Cubes 3.9.3 Viability Test on Dehydrated Fruit Cubes

3.10 Physiochemical Analysis 3.10.1 Moisture Content 3.10.2 Determination of pH Value 3.10.3 Determination of Soluble Solid Content (oBrix) 3.10.4 Determination of Water Activity Level (aw)

3.10 Storage of Dehydrated Fruit Cubes 3.11 Statistical Analysis

CHAPTER 4 RESULTS AND DISCUSSION 4.1 Growth of Lactobacillus acidophllus LA-5 (LA) and Lactobacillus casei

Le-OJ (LC) in three fruit juices 4.1.1 Growth of Lactobacillus acidophilus LA-5in papaya, banana and

pineapple juices for 48 Hours 4.1.2 Growth of Lactobacillus Casei Le-OJ in papaya, banana and

pineapple juices for 48 Hours 4.1.3 Statistical analysis on the growth of L. acidophilus and L. caseiin

there fruit juices 4.2 Immobilization of L. acidophilus and L. casei on fruit cubes with

different sizes 4.2.1 The immobilization of L. acidophilus and L. casei on papaya fruit

cubes 4.2.2 The immobilization of L. acidophilus and L. casei on banana fruit

cubes 4.2.3 The immobilization of L. acidophilus and L. casei on pineapple

fruit cubes 4.3 The survival of immobilized cultures during oven dehydration

viii

28 29 29 30 31 31 33 34 35 35

37 37 37 38 38 39 40 40 40 41 41 42 42 42 42 42 43 43 43 43 44 44

45

45

45

47

50

52

52

55

57

59

4.3.1 The survival of immobilized L acidophilusand L caseion oven dried fruit cubes dehydrated at 50°C and 60°C

4.3.2 The different mean value of immobilized cultures on fruit cubes during dehydration process

4.4 The survival of immobilized cultures during storage 4.4.1 The survival of immobilized cultures (L acidophilusand L casel)

on oven dehydrated and freeze dried papaya cubes (1.5 em3 and 1.0 em3

) during storage 4.4.2 The survival of immobilized cultures (L. acidophilus and L easel)

on oven dehydrated and freeze dried banana cubes (1.5 em3 and 1.0 em3

) during storage 4.4.3 The survival of immobilized cultures (L. acidophilus and L. easel)

on oven dehydrated and freeze dried pineapple cubes (1.5 em3

and 1.0 cm3) during storage

4.5 Rehydration of fruit cubes immobilized with probiotics

CHAPTER 5 CONCLUSION AND SUGGESTION

REFERENCES

APPENDIX

IX

59

61

62

62

65

66

68

74

76

91

Table 2.1: Table 2.2: Table 2.3: Table 2.4: Table 2.5: Table 2.6: Table 4.1:

Table 4.2:

Table 4.3:

UST OF TABLES

Prominent types of functional food lactobacilli used as probiotic cultures Bifidobacterium cultures used as probiotic cultures Some commercial examples of probiotic products Benefits of Probiotics The Top Five Commodity of Fruits in Malaysia 2007 Correlation between growth and pH value of cultures in fruits medium The mean of maximum viable count and time taken to achieve highest growth for Lactobacl1/us acidophilus and Lactobacillus caseiin three fruits juices respectively The survival of immobilized cultures after oven dehydration at 50°C and 60°C within 18 hours

x

Pages

9 11 12 13 16 34 47

50

62

LIST OF FIGURES

Pages

Figure 2.1: Awareness of consumers on health effects of various 10 functional ingredients

Figure 4.1: Growth of Lactobacillus acidophilus in papaya, pineapple 46 and banana medium respectively.

Figure 4.2: Growth of Lactobacillus caseiin papaya, pineapple and 49 banana medium respectively.

Figure 4.3: Effect of dimensions (1.0 em3 and 1.5 cm3) and 53

techniques of immersion prior to fermentation of medium (A) and after fermentation (8) towards the attachment of cultures (LA and LC) on papaya fruit cubes.

Figure 4.4: Effect of dimensions (1.0 cm3 and 1.5 cm3) and 56

techniques of immersion prior to fermentation of medium (A) and after fermentation (8) towards the attachment of cultures (LA and LC) on banana fruit cubes.

Rgure 4.5: Effect of dimensions (1.0 cm3 and 1.5 cm3) and 58

techniques of immersion prior to fermentation of medium (A) and after fermentation (8) towards the attachment of cultures (LA and LC) on pineapple fruit cubes.

Figure 4.6: The survival of L. acidophilus represented by (A) and L. 60 casei represented by (8) immobilized on fruit cubes sized 1.0 cm3 (using technique of immersion along fermentation) after oven dehydrated at 50De and 60De

Figure 4.7: Survival of immobilized LA and LC on papaya cubes after 64 dehydration (freeze dried or oven dried) and kept at -20De and room temperature (rt). Papaya cubes sized 1.5cm3 represented by (A), while papaya cubes sized 1.0 cm3 represented by (8)

Figure 4.8: Survival of immobilized LA and LC on banana cubes after 65 dehydration (freeze dried or oven dried) and kept at -20DC and room temperature (rt). Papaya cubes sized 1.5cm3 represented by (A), while papaya cubes sized 1.0 cm3 represented by (8)

Rgure 4.9: Survival of immobilized LA and LC on pineapple cubes 67 after dehydration (freeze dried or oven dried) and kept at -20De and room temperature (rt). Papaya cubes sized 1.5cm3 represented by (A), while papaya cubes sized 1.0 cm3 represented by (8)

Rgure 4.10: Rehydration ratios of fruit cubes sized 1.0 cm3 with (A) 69 represent freeze dried fruit cubes and (8) represent oven dried fruit cubes. Wt shows weight of rehydrated fruit cubes at speCific time while Wo shows dehydrated fruit cubes

Figure 4.11: Rehydration ratios of fruit cubes sized 1.5 cm3 with (A) 72

xi

represent freeze dried fruit cubes and (8) represent oven dried fruit cubes. Wt shows weight of rehydrated fruit cubes at specific time while Wo shows dehydrated fruit cubes

XlI

cfu/ml Cm cm3

g kg log cfu g-l ml Ph aw % °Brix °C CAGR ANOVA FAO LA-5 lC-Ol WHO

UST OF SYMBOLS

viable cell number centrimeter centrimeter cubes gram kilogram The log value of viable cell number mililiter Acidity measured in pH Water activity percentage degree Brix degree Celcius compound annual growth rate Analysis Of Varians Food and Agriculture Organization of the United Nations Lactobacillus addophilus 5 Lactobacillus casei 01 World Health Organization

xiii

UST OF APPENDIX

Pages

Appendix Al: Correlation between growth of LA and pH of Banana 91 medium

Appendix A2: Correlation between growth of LC and pH of Banana 92 medium

Appendix A3: Correlation between growth of LA and pH of papaya 93 medium

Appendix A4: Correlation between growth of LC and pH of papaya 94 medium

Appendix AS: Correlation between growth of LA and pH of pineapple 95 medium

Appendix A6: Correlation between growth of LC and pH of pineapple 96 medium

Appendix A7: t-test of LA and LC in papaya medium 97 Appendix A8: t-test of LA and LC in Banana medium 98 Appendix A9: t-test of LA and LC in pineapple medium 99 Appendix Bl: pH reading of immobilized fruit cubes (papaya) 100 Appendix B2: pH reading of immobilized fruit cubes (banana) 101 Appendix B3: pH reading of immobilized fruit cubes (pineapple) 102 Appendix 84: Pictures of fruit cubes before and after dehydration 103 Appendix Cl: Table of moisture content and water activity during 106

dehydration Appendix C2: One way Anova analysis for viability of culture on 108

immobilized fruit cubes during dehydration Appendix 01: Moisture content of dehydrated papaya, banana and 109

pineapple cubes during storage Appendix 02: Water activity of dehydrated papaya, banana and 121

pineapple cubes during storage Appendix 03: pH value of dehydrated papaya, banana and pineapple 133

cubes during storage Appendix 04: Graph of the immobilized cultures survival with 145

different technique of immersion or fermentation

xiv

CHAPTER 1

INTRODUCTION

The increasing interest of consumers in their personal health has lead to the

development of functional foods (Doleyres & lacroix, 2005). Functional foods are

foods that contain some health-promoting components beyond traditional nutrients

(Shah, 2001). lactic acid bacteria are normal components of the intestinal microflora

in both humans and animals which have been associated with various health­

promoting properties (Pereira et aI., 2003). This benefit has lead to the development

of functional food products containing lactic acid bacteria for human consumption.

These beneficial lactic acid bacteria or sometimes known as probiotic can be found in

infant foods, cultured milks, and various pharmaceutical preparations (Sanders,

1999).

Probiotics are well-defined bacterial types administrated to the host in

sufficient numbers to confer defined and proven physiological benefits (Reid, 2007).

In this new era, food manufacturers are trying to include probiotic strains in foods

and beverages which are part of a normal diet to provide health defense while

enjoying meals and to differentiate such functional products from concentrated

probiotic preparations available as capsules, powders or liquids (lavermicocca et aI.,

2005). lactobacillus acidophil us and lactobacillus casei are the example of lactic acd

bacteria which can be found worldwide in a variety of products, including

conventional food products and dietary supplements (Reid, 2007).

In order to exert probiotics beneficial effects In the host, it is generally

accepted that probiotic bacteria must be alive in product at the time of consumption

and also capable of reaching the large intestine in high enough quantities to facilitate

colonization or even proliferation (Shah, 2000) and in general a minimum level of

more than 106 viable probiotic bacteria per mililitre or gram of food product is

accepted to beneficial human health when consumed (Ouwehand & Salminen, 1998).

There is growing scientific evidence that maintenance of healthy gut microbiota can

provide several health benefits for the host that probiotics might contribute (Saxelin

et aI., 2005).

A number of genere of bacteria and yeast are used as probiotics, including

Lactobacillus, Leuconostoc, Pedlococcus, Bifidobacterium and Enterococcus, but the

main species believed to have probiotics properties are Lactobacillus acidophilus,

Bifidobacter/um spp. and Lactobacillus casei (Shah, 2004). Lactobacillus acidophilus

is the most commonly used organism as dietary supplement and is often added to

fermented milks because of its probiotic effects (Curry & Cow, 2003; Olson & Aryana,

2007). However, Lactobacillus acidophilus tends to grow slowly in milk and do not

survive well in fermented milk due to the low pH, and it is difficult to maintain large

numbers in the product (Shah, 2007). Lactobacillus casel is another example of

probiotic that present in fermented dairy products and has beneficial properties for

human health (Oozeer et aI., 2002). From the human trial that was carried out to

assess ileal and faecal survival of the probiotic bacterium, it was concluded that

Lactobacillus casei can survive transit through human gut (Oozeer et aI., 2006).

Probiotics play an important role in antimicrobial activity and gastrointestinal

infections and for instance, Lactobacillus acidophilus produces various bacteriocins

and antibacterial substances such as Lactocidin, Acidolin, Acidophilin, Lactaclum-B

and inhibitory protein which inhibit several pantogenic bacteria (Shah, 2000).

Probiotics are claimed to shorten duration of rotavirus diarrhoea in children as well

(Saavedra et aI., 1994), for example there is a pediatric beverage containing mix of

Bifidobacterium anlmalis, Lactobacillus acidophilus and Lactobacillus reutri produced

to prevent rotavirus diarrhoea in children (Guandalini et aI., 2000). The most widely

accepted health benefit of probiotic organism is the improvement of lactose

metabolism (Shah, 2000). Others benefits are such as antimutagenic properties that

provide efficient inhibition of mutagens (Shah, 2007). Probiotic organisms are also

popular with their ability to remove sources of procarcinogens (Singh et aI., 1997).

These beneficial organisms also de-conjugated bile salts in human body and as a

result reduce the cholesterol in body (Klaver & Meer, 1993).

Probiotic cells entrapment in food grade porous matrix has been the most

widely used immobilization technique to produce solid food with probiotic

(Champagne et al, 1994). Koukoutas (2000) previous studies shown that apple

2

pieces are promising carriers for probiotic bacteria and may be used in the

production of probiotic fermented milk or other food products, as well as In the

prolongation of their shelf-life. Fruits are chosen as food matrix for Lactobacillus

immobilization due to their food-grade purity, cheap and abundant in nature while

the immobilization technique is simple and easy and showed high operational

stability (Kourkoutas et al, 2004). It is strongly believed that future clinical tests will

ensure the beneficial effects of fruit based problotics. Freeze-dried apple supported

Lactobacillus casei biocatalyst could be added to various solid foods for examples

breakfast cereals, used in baking and etc. to provide probiotic properties (Kourkoutas

et al, 2006).

The ability of microorganisms to grow and survive depends largely on their

capacity to adapt to changing environments (Doleyres & LacrOix, 2005). Probiotic

must exhibit high survival rates in downstream processes such as centrifugation and

dehydration, also in food products during storage (Saarela, 2008). Downstream

processes for example dehydration lead to the exposure of live probiotic bacteria to a

variety of stresses, such as heat, cold, oxygen and osmotic stresses, leading to

impaired functionality and loss of vialbility during drying and storage (Meng et al,

2008). Viability and survival of probiotic are important to make sure the probiotic

reaches the human intestinal and bring health benefits (Saarela, 2008). lactobacilli

especially Lactobacillus acidophil/us and Lactobacillus casel groups are generally

considered to be more resistant to acidic environments than blfidobacteria

(Champagne & Gardner, 2005). A development of more efficient technologies could

lead to greater product efficacy and strain diversification with the development of

technologically unsuitable strains into products (Lacroix & Yildirim, 2007).

There are strong demands for new technologies that enable high probiotic

cells yield at large scale incorporated into inexpensive abundant non dairy food

matrixes without losing viability and functionality. Since the application of probiotic

cultures in non-dairy faces a great challenges this research of the growth and

survival of probiotic cells immobilized on dehydrated fruit cubes is being carried out

with the following specific objectives:

1. To identify the growth of Lactobacillus CiJsei and Lactobacillus acidophilus in

papaya, banana and pineapple fruit juices as immobilization medium.

3

2. To determine the survival rate of Lactobacillus casei and Ladobacillus

acidophilus immobilized on papaya, banana and pineapple fruits cubes.

3. To enumerate the probiotic culture cells immobilized on different sizes of fruit

cubes.

4. To investigate the survival rate of immobilized probiotic cult\Jres after

dehydration processes and during the storage of 56 days.

4

CHAPTER 2

LITERATURE REVIEW

2.1 Roles of Functional Foods to Public Health

Food industry companies have rather high expectations in food products that meet

the consumers' demand for a healthy life style. In this context Functional Food playa

specific important role (Menrad, 2003). Formulated to be significant sources of

specific nutrients or other components of nutritional significance, functional foods

have the potential to make a major impact on the food supply and public health. The

potential success of functional foods in developed countries can be attributed to two

trends. Increasingly affluent and ageing populations have become more concerned

with protecting their health through diet (Menrad, 2003). At the same time, longer

working hours and interest in fulfilling leisure activities have left younger individuals

finding it difficult to purchase and prepare foods to meet dietary recommendations

(Mogelonsky, 1999).

At the same time, on a global scale, climate change, growing world

populations, and clear evidence of the depletion or damage to critical resources

makes it imperative to take action to sustain and develop the capacity of agricultural

and manufacturing systems to continue to provide food, the most basic of human

needs. Despite major advances in technology leading to more efficient food

production, millions of people are hungry or severely undernourished, while an

almost comparable number are dangerously overnourished (Aiking & Boer, 2004).

Food manufacturers awareness of the potential commercial opportunity, and the

deregulation of health claims (Katan, 1999) and in some cases, lack of regulation,

have contributed to growth of this sector of the food industry (Menrad, 2003).

Starting 1980s, enormous progress has been made in establishing the

scientific basis for functional food development in health, nutrition and food

processing (Diplock et al., 1999 & Mermelstein, 2002). Functional foods are founded

on the key premise that, compared to conventional foods, they help to ens\Jre overall

good health and to prevent or manage specific conditions in a convenient way

through daily diet (Sloan, 2000).

The introductions of foods with additional health value which benefits the

public offer interesting growth opportunities for the food industry (Kleef et aI., 2005).

Most early developments of functional foods were those of fortified with vitamins and

minerals such as vitamin C, vitamin E, folic acid, zinc, iron, and calcium (Sloan,

2000). Subsequently, the focus shifted to foods fortified with various micronutrients

such as omega-3 fatty acid, phytosterol, and soluble fiber to promote good health or

to prevent diseases such as cancers (Sloan, 2002). More recently, food companies

have taken further steps to develop food products that offer multiple health benefits

in a single food (Sloan, 2004).

In general, functional products are 'add good to your life' for improve the

regular stomach and colon functions or 'improve children'S life' by supporting their

learning capability and behaviour. It is difficult, however to find good biomarkers for

cognitive, behavioural and psychological functions. Whereas, other groups of

functional food is designed for reducing an existing health risk problem such as high

cholesterol or high blood pressure. Functional foods also consist of products, which

'makes your life easier' for example the development of lactose-free and gluten-free

products (Makinen-Aakula, 2006).

2.1.1 The Development of Functional Foods

The globalization of food technologies in this new decade has lead to the idea that

foods are not only intended to satisfy hunger and to provide necessary nutrients for

humans but also to prevent nutrition-related diseases and improve physical and

mental well-being of the consumers (Menrad, 2003). Functional foods are gaining in

popularity, big-name grocery product manufacturers like Kraft are making huge

research and development investments in bringing these nutritionally enhanced

foods to market as customer demand for healthier enhanced foods Is surging

(Adams, 2004).

6

The term functional food itself was first used in Japan, in the 1980s, for food

products fortified with special constituents that possess advantageous physiological

effects (Hardy, 2000; Kwak & Jukes, 2001; Stanton et al., 2005). Then the concept

of functional food was promoted in 1984 by Japanese scientists who studied the

relationships between nutrition, sensory satisfaction, fortification and modulation of

physiological systems. The Japan Ministry of Health introduced rules for approval of a

speCific health-related food category called FOSHU ( Food for Specified Health Uses)

in 1991 which included the establishment of specific health claims for this type of

food (Burdock et al., 2006; Kwak & Jukes, 2001; Menrad, 2003 and Roberfroid,

2000).

In total, more than 1700 functional food products have been launched in

Japan between 1988 and 1998 with an estimated turnover of around 14 billion US

dollar in 1999 (Menrad, 2003). The market was estimated to be 5 billion US dollar in

2003 (Side, 20060re than 500 products were labelled as FOSHU in 2005 (Fern, 2007

& Side, 2006). There is no doubt that the Japanese interest in functional foods has

also brought awareness for the need of such products in places like Europe and the

United States. Experts in these countries realized that besides being able to lower

the cost of healthcare of the aging population, functional food might also give a

commercial potential for the food industry (Siro et a/., 2008).

The global market of functional food is estimated to at least 33 billion US

dollar (Hilliam, 2000). Other experts like Sloan (2000) and Sloan (2002) has

reckoned the global functional food market to be 47.6 billion US dollar, being the

United States the largest market segment, followed by Europe and Japan. Some

estimations report even more global market value of nearly 61 billion US dollar

(Benkouider, 2004). The three dominant markets contribute over 90% of the total

sales (Benkouider, 2005).

Not only food manufacturers, but also the pharmaceutical industry has

become interested in this field. In consequence of this has led to the so-called grey

area which describes the overlapping of the interests of food and pharmaceutical

industries (Farr, 1997; Kotilainen et a/., 2006 & Mark-Herbert, 2004). This latter is

represented by several companies such as Novartis Consumer Health, Glaxo

Smith Kline, Johnson & Johnson or Abbott Laboratories. One important motivation for

7

such companies to invest in functional food is the shorter development times and

lower product development costs compared to pharmaceutical products. In addition,

these companies have intensive experience in organizing clinical trials to substantiate

health claims of a specific product. Against the above-mentioned, pharmaceutical

companies generally failed to gain foothold in the functional foods market due to the

incompetence of developing and marketing a high-quality food product (Bech-Larsen

& Scholderer, 2007).

A third group of functional food producers are companies specialized in a

particular product category, which mostly belong to the market leaders on a national

level, for example Molkerei Alois Muller (,'proCulr' dairy products), Eckes (ACE drinks)

or Becker Fruchtsafte (ACE fruit juice) in Germany (Siro, 2008). There is a limited

number of small and medium-sized food companies (SMEs) active in the functional

food market as well. These companies mostly produce functional products for market

niches or offer "me-too" products following the pioneering products of the

multinational companies. Often these products can survive only for a rather short

time period for example only up to two years. In general, SMEs lack the know-how

and resources for own intensive Research and development activities and cannot

afford to spend high sums in specific information or advertising activities necessary

to open a speCific segment of the functional food market as pioneering company

(Menrad, 2003).

Food retail companies are increasingly starting to introduce private label

brands especially in the relatively mature markets of functional dairy products. In

Germany, for example, this relates in particular to food discounters like Aldi, Udl and

Penny, which have launched probiotic and prebiotic dairy products in recent years

(Menrad, 2003). Functional foods have been developed in virtually all food

categories. From a product point of view, the functional property can be included in

numerous different ways as it can be seen in Table 2.1.

8

Table 2.1: Prominent types of functional food

Type of functional food Definition Example

Fortified product A food fortified additional nutrients

with Fruit juices fortified with vitamin C

Enriched products

Altered products

Enhanced commodities

A food with added new nutrients or components not normally found in a particular food A food from which a deleterious component has been removed, reduced or replaced with another substance with beneficial effects A food in which one of the components has been naturally enhanced through special growing conditions, new feed composition, genetic manipulation, or otherwise

Sources: Kotilainen et aI., 2006 & Spence, 2006.

Margarine with plant sterol ester, probiotics, prebiotics

Fibers as fat releasers in meat or ice cream products

Eggs with increased omega-3 content achieved by altered chicken feed

Functional food products are not homogeneously scattered over all segments of the

food and drink market and consumer health concerns and product preferences may

vary between markets. These products have been mainly launched in the dairy-,

confectionery-, soft-drinks-, bakery- and baby-food market (Kotilainen et aI., 2006 &

Menrad, 2003).

The future market development is influenced by the degree of familiarity and

acceptance of Functional Food as well. According to surveys in different European

countries consumers often do not know the term Functional Food but show a rather

high agreement to the concept. In the United Kingdom, France and Germany, up to

75% of the consumers have not heard the term 'Functional Food', but more than

50% of them agree to fortify functional ingredients in specific food products (Hilliam,

1999). Thereby the acceptance to a speCific functional ingredient is linked to the

consumer's knowledge of the health effects of specific ingredients. Therefore,

functional ingredients which are in the mind of consumers for a relatively long period

of time for example vitamins, fiber, calcium and iron achieve considerably higher

rates of consumer acceptance than ingredients which are used for a short period of

9

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