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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[itusi pengajian tinggi.
4.
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Si1a tandakan (/ )SULIT
TERHAD
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,TIDAK TERHAD~ (TAND~PENULIS)
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(Mengandungi maklumat TERHAD yang telah ditenhlkakan oleh organisasilbadan di mana peny lidikan dijalankan)
Disahkan oleh
(TA iDATANGAN PU TAKAWAN)
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Jika tesis ini SULIT atau TERHAD, si1a lampiran surat daripada pihak berkuasa/organsasi berkenaan dengan menyatakan seka!i sebab dan tempoh tesis ini perlu dikelaskan sebagai SULITdanTE~AD. . . . . /
*
Tesls dlmaksudkan sebagal teslS bagl ~azah Doktor Falsafah dan Sarjana secara penyeJiaikan, atau diserta?1 bagi pengajian secara kerja kursus dan penyelidikan, atau Laporan Projek 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.
ABSTRAK
PERnJHBUHAN DAN KEUPAYAAN UNnJK HIDUP PROBIOTIK YANG DINYAHHOBIUSASIKAN PADA KIUB BUAH-BUAHAN KERING
Dalam penyelidikan ini. kesan penggunaan cara fermentasi yang berlalnan, buah- buahan, 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
mempunyaipertumbuhan 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
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
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 -
20D
e
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
represent freeze dried fruit cubes and (8) represent oven dried fruit cubes.
W
t shows weight of rehydrated fruit cubes at specific time while Wo shows dehydrated fruit cubesXlI
cfu/ml Cm cm3 g
kg
log cfu g-l ml
Ph
a
w%
°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
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 (Kourkoutaset
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.
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 HealthFood 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 overallgood 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 (Burdocket 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 forsuch 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
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