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Food Research International 41 (2008) 111–123 www.elsevier.com/locate/foodres
Trends in non-dairy probiotic b
? Flavera C. Prado a, Jose L. Parada a, Ashok Pandey b, Carlos R. Soccol
Laboratory of Biotechnology Process, Department of Chemical Engineering, Federal University of Parana, 81531-970 Curitiba, PR, Brazil b Biotechnology Division, Regional Research Laboratory, CSIR, 695 019 Trivandrum, India Received 30 May 2007; accepted 24 October 2007
Abstract In recent times, there has been an increased interest to adapt healthy diets, which help in preventing diseases, and as a consequence, the study and development of new functional foods has gained much importance. Food additives as probiotics and prebiotics may exert positive e?ects on the composition of gut microbiota and are subject of intensive research. The allergy to dairy products a?ect negatively some persons. Lactose intolerance and the cholesterol content are two major drawbacks related to the fermented dairy products. Traditions and economic reasons that limit the use of dairy fermented products in some developing countries promote the idea of reduction of milk components as vehicles for the probiotic agents. At present, some non-dairy probiotic beverages are being commercialized and are discussed in this review. Probably, beverages such as fruit and vegetable juices would be the next food category where the healthy probiotic bacteria will make their mark. ? 2007 Elsevier Ltd. All rights reserved.
Keywords: Functional foods; Probiotics; Prebiotics; Fermented milks; Non-dairy beverages
Contents 1. 2. 3. 4. 5. Introduction . . . . . . . . . . . . . . . . . . . . . . Probiotics as functional foods . . . . . . . . . . Achievements of dairy probiotic beverages . Non-dairy probiotic beverages . . . . . . . . . Perspectives of probiotic beverages . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 111 113 115 117 120 120
1. Introduction The development of probiotics in the last two decades has signaled an important advance in the food industry. The number of scienti?c publications on probiotics has increased a lot stimulated by factors as exciting scienti?c and clinical ?ndings using well-documented probiotic
Corresponding author. Tel.: +55 41 33613191; fax: +55 41 33613674. E-mail address: email@example.com (C.R. Soccol).
organisms. Some concerns over the limitations and side e?ects of the pharmaceutical agents and consumers demand for the natural products have been discussed by Reid (2006). Probiotic is a relatively new word meaning ‘‘for life’’ and it is generally used to name the bacteria associated with the bene?cial e?ects for the humans and animals. The term probiotic was technically de?ned by an Expert Committee as ‘‘live microorganisms which upon ingestion in certain numbers exert health bene?ts beyond inherent general
0963-9969/$ - see front matter ? 2007 Elsevier Ltd. All rights reserved. doi:10.1016/j.foodres.2007.10.010
112 Table 1 Probiotic microorganisms Lactobacillus species Lb. acidophilus Lb. amylovorus Lb. brevis Lb. casei Lb. casei sp. rhamnosus Lb. crispatus Lb. delbrueckii sp. bulgaricus Lb. fermentum Lb. gasseri Lb. helveticus Lb. johnsonii Lb. lactis Lb. paracasei Lb. plantarum Lb. reuteri Bi?dobacterium species Bf. Bf. Bf. Bf. Bf. adolescentis animalis breve bi?dum infantis
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Others Bacillus cereus Clostridium botyricum Enterococcus faecalis Enterococcus faecium Escherichia coli Lactococcus lactis sp. cremoriss Lactococcus lactis sp. lactis Leuconostoc mesenteroides sp. dextranicum Pediococcus acidilactici Propionibacterium freudenreichii Saccharomyces boulardii Streptococcus salivarius sp. thermophilus
Bf. lactis Bf. longum
Cited by Collins, Thornton and O’Sullivan (1998), Gorbach (2002), Holzapfel and Schillinger (2002) and O’Sullivan et al. (1992).
nutrition’’ (FAO/WHO, 2001; Guarner & Schaafsma, 1998). This means that the microorganisms must be alive and present in high numbers, generally more than 109 cells per daily ingested dose. Each product should indicate the minimum daily amount required for it to confer speci?c health bene?t(s). The probiotic microorganisms consist mostly of strains of the genera Lactobacillus and Bi?dobac-
terium, but not exclusively (Table 1). These types of lactic acid bacteria have been used since recorded history in the production of fermented dairy products (FAO/WHO, 2001; Gorbach, 2002). No pathogenic, or virulence properties have been found for the lactobacilli, bi?dobacteria, or lactococci (Aguirre and Collins, 1993). But, in terms of safety of the probiotics, FAO/WHO Expert Consultation believes that principles and practical criteria need to be generated to provide the guidelines as to how any given potential probiotic microorganism can be tested and proven to have a low risk of inducing, or being associated with the etiology of some disease, versus conferring a signi?cant health bene?t when administered to humans (FAO/WHO, 2001). There are some ideal properties of the probiotic strains, which would bene?t the human health and could be used in probiotic industry: resistance to acid and bile; attachment to the human epithelial cells; colonization in the human intestine; production of antimicrobial substances, called bacteriocins (Jack, Tagg, & Ray, 1995); good growth characteristics and bene?cial e?ects on the human health, such as presented in Fig. 1. One of the most important characteristics of a probiotic strain is that it must be non-pathogenic and GRAS – Generally Regarded As Safe (Collins, Thornton, & O’Sullivan, 1998; Gorbach, 2002; Havenaar & Huis In’t Veld, 1992). The probiotic must also present some desirable characteristics, such as low cost, maintain its viability during the processing and storage, facility of the application in the products, resistance to the physicochemical processing of the food. There are di?erent degrees of evidence supporting the health e?ects of the probiotics (FAO/WHO, 2001). These
Stimulation of the immune response
Treatment and prevention of acute diarrhoea by rotaviruses
Increase of the lactose tolerance and digestion
Restoration of the normal intestinal microflora after antibiotic therapy
Positive influence in the intestinal microflora
Production of the B vitamins (folic acid)
Reduction of intestinal pH
Reduction of ammonia and other toxic compounds
Improvement of the intestinal functioning
Fig. 1. Probiotic bene?cial e?ects on human health. Adapted from Gibson & Roberfroid (1995).
F.C. Prado et al. / Food Research International 41 (2008) 111–123 Table 2 Examples of probiotic strains used in prevention and treatment of some diseases Disorders Diarrhea caused by pathogenic bacteria and viruses Strains Lb. rhamnosus GG, Lb. casei Bf. lactis BB-12, Bf. bi?dum Sc. thermophilus Clinical outcomes Prevention and treatment of acute diarrhea caused by rotaviruses in children References
Helicobacter pylori infection and complications
Lb. johnsonii La1, Lb. salivarius, Lb. acidophilus LB
In?ammatory diseases and bowel syndromes Cancer in gastrointestinal tract
Lb. rhamnosus GG
Lb. rhamnosus GG, Lb. rhamnosus LC-705, Lb. casei Shirota, Lb. acidophilus LA-2, Bi?dobacterium sp., Propionibacterium sp. Lb. casei Shirota, Lb. rhamnosus HN001, Lb. acidophilus HN017, Bf. Lactis HN019
Inhibition of the pathogen growth and decrease urease enzyme activity necessary for the pathogen to remain in the acidic environment of the stomach Remediation in in?ammatory conditions through modulation of the gastrointestinal micro?ora Prevention or delay of the onset of certain cancers
Guarino, Berni, Spagnuolo, Albano, and Di Benedetto (1997), Guandalini et al. (2000), Isolauri, Juntunen, Rautanen, Sillanaukee, and Koivula (1991), Majamaa, Isolauri, Saxelin, and Vesikari (1995), Perdone, Bernabeu, Postaire, Bouley, and Reinert (1999), Saavedra, Bauman, Oung, Perman, and Yolken (1994), Shornikova, Isolauri, Burkanova, Lukovnikova, and Vesikari (1997) and Szajewska, Kotowska, Mrukowicz, Armanska, and Mikolajczyk (2001) Aiba, Suzuki, Kabir, Takagi, and Koga (1998), Coconnier, ? Lievin, Hemery, and Servin (1998), Luo and Grayson (1995), Michetti et al. (1999) and Midolo, Lambert, Hull, Kabir et al. (1997) Giochetti et al. (2000), Gupta, Andrew, Kirschner, and Guandalini (2000) and Shanahan (2000) Aso et al. (1995), El-Nezami, Mykkanen, Kankaanpaa, ¨ ¨¨ Salminen, and Ahokas (2000), Hosada, Hashimoto, He, Morita, and Hosono (1996) and Oatley, Rarick, Ji, and Linz (2000)
Enhancement of immune parameters
Lb. rhamnosus GG, Bf. lactis BB-12 Lactobacillus sp. Lb. acidophilus, Lb. rhamnosus GG
Prevention of allergic diseases onset Prevention and therapy of ischemic heart syndromes Eradication of vaginitis through restoration of dominated vaginal ?ora Lower risk of urinary tract infections through restoration of dominated vaginal ?ora
Cardiovascular disease Bacterial and yeast vaginitis
Urinary tract infections
Lactobacillus GR-1, Lactobacillus B-54, Lactobacillus RC-14
Arunachalam, Gill, and Chandra (2000), Donnet-Hughes, Rochat, Serrant, Aeschlimann, and Schi?rin (1999), Gill, Rutherfurd, Prasad, and Gopal (2000), Gill, Cross, Rutherfurd, and Gopal (2001), Matsuzaki and Chin (2000), Sheih, Chiang, Wang, Chuh, and Gill (2001) and Perdigon, Vintini, Alvarez, Medina, and Medici (1999) Isolauri, Arvola, Sutas, Moilanen, and Salminen (2000), Kalliomaki et al. (2001) and Majamaa and Isolauri (1996, 1997) De Roos and Katan (2000) and Oxman, Shapira, Klein, Avazov, and Rabinowitz (2001) Hilton, Isenberg, Alperstein, France, and Borenstein (1992), Hilton, Rindos, and Isenberg (1995), Reid et al. (2001a), Reid, Beuerman, Heinemann, and Bruce (2001b) and Sieber and Dietz (1998) Reid, Bruce, and Taylor (1995) and Reid et al. (2001b)
Adapted from FAO/WHO (2001).
have been established by the scienti?c testing in the humans, performed by the legitimate research groups and published in peer-reviewed biomedical journals (Gorbach, 2002). Several examples are shown in Table 2, with some speci?c strains and clinical outcomes. Some of them are in the market (Table 3) and make good pro?ts for the companies. 2. Probiotics as functional foods The concern in consuming innocuous foods free of pathogenic microorganisms, or their toxins, or containing the microorganisms, which can alter their chemical components, has emerged in the developing countries. Moreover, they must not be contaminated with any undesirable substances or, at least, they must ful?ll the maximum limits
of the residues allowed after exhausting studies demonstrating the risks. On the other hand, the interest of the consumers to ingest diets to maintain the health and prevent the degenerative, or chronic diseases such as diabetes, cancer, hypertension and others, has been in?uenced more and more, by the current of favorable opinions towards the consumption of the natural foods and limit processed foods. Since the 1980s, this and other factors have conduced to the study and development of the so-called functional foods. The interest in developing these products is rising day-by-day, driven by the market potential for the foods and beverages that can improve the health and well-being of the consumers (Hilliam, 2000). The functional foods have also been termed as medicinal foods, nutraceuticals, prescriptive foods, therapeutic foods, super-foods, designer foods, foodiceuticals and medifoods
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Table 3 Probiotic lactic acid bacteria marketed worldwide Strains Lactobacillus Lactobacillus Lactobacillus Lactobacillus casei Shirota crispatus CTV05 reuteri MM53 casei F19 Origin Yakult, Japan Gynelogix, USA BioGaia, Sweden Arla Foods, Denmark/ Sweden Danisco, France Valio, Finland
Bi?dobacterium lactis HN019 Lactobacillus rhamnosus GG Propionibacterium freudenreichii ssp. shermanii JS Lactobacillus acidophilus NCFM Lactobacillus acidophilus NCFB 1748 Lactobacillus johnsonii LA1 (NCC 533) Lactobacillus acidophilus LA10 (NCC 90) Lactobacillus fermentum RC-14 Lactobacillus rhamnosus GR-1 Lactobacillus casei DN-114 001 Bi?dobacterium animalis DN-173 010 Lactobacillus plantarum 299v Lactobacillus rhamnosus 271 Lactobacillus casei CRL 431 Bi?dobacterium lactis BB-12 Lactobacillus acidophilus LA-5 Lactobacillus bulgaricus LBY27 Streptococcus thermophilus STY-31
Rhodia, USA ? Nestle, Switzerland Urex, Canada Danone, France Probi AB, Sweden Chr. Hansen, USA
(Finley, 1996). Because of the good amount of information available currently, it is convenient to de?ne the concept of the functional foods and di?erentiate them from other terms such as nutraceutics, or forti?ed and special formulas. The functional food concept was developed during the 1980s in Japan and it gained legal status in 1991, being described as FOSHU, or ‘‘Foods for Speci?ed Health Use’’. The country has formulated a speci?c regulatory process for the functional food approval. And it has more than 100 products with FOSHU license (Sanders, 1998). Functional foods, as they are known in Japan, do not exist in either Europe, or the United States because in these countries, no regulation, or policy statements exist, specifically for them (Stanton et al., 2001). Legislation in Brazil is recent. Functional foods are regulated by ANVISA – National Agency of Sanitary Vigilance through Governmental Decree n. 15 e Resolutions n.16, 18 and 19 dated April 1999 (Brasil, 1999a, 1999b, 1999c, 1999d; Oliveira, Sivieri, Alegro, & Saad, 2002). The norms that regulate functional foods must consider the following aspects in relation to the probiotics: the preparation should remain viable in large-scale production; it should be stable and viable during the storage and use; the preparation should be able to survive in the intestinal ecosystem; and, the host must get bene?ts to lodge probiotics. Functional foods are de?ned as the foods that in addition to nutrients, supply the organism with components that contribute to cure the diseases, or to reduce the risk of developing them. They are foods like the conventional ones, which produce proved physiological e?ects and/or
reduce the risk of developing chronic diseases and could be similar in physical appearance to the conventional food, consumed as part of the daily dietary, but able to produce proved metabolic, or physiological e?ects, useful in the maintenance of a good physical and mental health and to able to help in the reduction of the risk of degenerative chronic diseases, besides to contribute to its basic nutritional functions. They could be foods with functional compounds, which have physiological e?ects on the consumer, besides their basic nutritive value. The main aim of these products is to prevent chronic diseases and to maintain the natural balance of vitamins and electrolytes of the body. According to the International Life Science Institute (ILSI), a food can be considered functional if it can demonstrate satisfactorily that it has a bene?cial e?ect on one, or several speci?c functions in the organism, beyond the normal nutritional e?ects that improve the state of health and well-being, or reduce the risk of a disease. It is necessary to know what is meant by ‘‘beyond the normal nutritional e?ects’’. The feeding de?ned in the recommended daily dose can contain, in addition to usual foods, restored foods (restoration of the initial content of vitamins and minerals) and enriched foods; the dietetic foods respond to the specify necessities of some consumers. The functional foods are beyond these basic necessities. However, they must continue being foods and must be consumed in compatible amounts with a balanced and diversi?ed normal feeding. A functional food can be a natural one; or foods added of components; or foods of which component have been eliminated by means of technological or biotechnological procedures. It also can be a food, in which the nature of one or more of its component has been modi?ed, or a food in which the bio-availability of one or more of its components has been modi?ed, or any combination of the possibilities as above. The development of functional foods on the part of food industries and the clinical tests that are made to evaluate their e?ects will help the national and international sanitary authorities to establish clear criteria to classify this type of foods. Nutraceuticals are de?ned as the foods manufactured from common foods and sold in the form of prepared like pills, capsules, powder, syrups, drinks, etc. They are medicinal formula, which are not associated generally with the foods and can exert a bene?cial physiological e?ect, or assure protection against the chronic diseases. Fig. 2 shows a graphical representation of nutraceutical foods. Functional foods, design foods and nutraceuticals are expressions that can be used indi?erently to talk foods, or isolated ingredients that provide determined non-nutritional bene?cial physiological e?ects that can improve the health. The foods are composed by thousands of biologically active constituents that can have a bene?cial e?ect on the health. The ?rst generation of functional foods involved supplementation of the constituents such as calcium, or vitamins, for their recognized health attributes (Ziemer & Gibson, 1998). Fermented dairy products can be included in the cat-
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egory of functional foods because of their content of calcium (which can reduce the osteoporosis, hypertension and colon cancer) and other health-enhancing components. In the recent decades, there is a greater interest in the potential bene?cial e?ects of the fermented milk on the health, resulting in the increase of the available variety and amount consumed around the world. Dairy products have been used traditionally like vehicle for the probiotic bacteria in humans. Lactic acid bacteria are the more important group of microorganisms used in the fermented milk elaboration and much of them are considered probiotics. Fermented milks are result of the metabolism of lactic acid bacteria that grow in milk. Its excellent nutritional quality can be attributed mainly to the milk, which o?ers an important source of calcium, proteins, phosphorus and ribo?avin. The additional bene?ts on the health are result of the fermentation process, which supplies fermented milk with a large number of microorganisms and products of fermentation. Fermented milks have probiotic e?ect because their consumption takes to the ingestion of high amounts of live bacteria that exert bene?ts in the balance of the intestinal micro?ora and the health, beyond the basic nutrition. Probiotic dairy foods containing healthpromoting bacteria are an important segment of the functional-food market (Batish & Grover, 2004). It represents a strong area in the growth within the group of functional foods and intense investigation to develop the dairy products with the probiotic microorganisms is in active progress. In fact, the concept of functional foods has become more directed towards food additives that may exert a positive e?ect on the gut microbiota composition. This has largely concentrated on the probiotics, but more recently interest in prebiotics has increased (Ziemer & Gibson, 1998). Prebiotics are non-digestible food ingredients that bene?cially a?ect the host by selectively stimulating the growth and/or activity of one, or a limited number of bacteria in the colon. This de?nition overlaps with the de?nition of dietary ?ber, with the exception of its selectivity for certain species (Gibson & Roberfroid, 1995; Schrezenmeir & De Vrese, 2001). Food ingredients classi?ed as prebiotics must not be hydrolyzed or absorbed in the upper gastrointestinal tract, need to be a selective substrate for one, or a limited number of colonic bacteria, must alter the microbiota in the colon to a healthier composition and should induce luminal, or systematic e?ects that are bene?cial to the host health (Gibson & Roberfroid, 1995). Peptides, proteins and lipids contain prebiotics characteristics, but some carbohydrates have received the most attention, including lactulose, inulin, and a range of oligosaccharides that supply a source of fermentable carbohydrate for the bene?cial bacteria in the colon (Crittenden, 1999; Ziemer & Gibson, 1998). The inulin and oligofructose are carbohydrates stored in the plants, presented in the foods as garlic, bananas, cereals, onions, and in high amounts in the root of the chicory. Oligofructose is a natural component of inulin (De Bondt, 2003). Symbiotics are
mixtures of the probiotics and prebiotics that bene?t the host by improving the survival and implantation of the selected microbial supplements. Because of the nutritional bene?ts associated with micro?ora management approaches, foods are the main vehicle for pro-, pre- and symbiotics (Gibson & Roberfroid, 1995; Ziemer & Gibson, 1998). 3. Achievements of dairy probiotic beverages Fermented beverages make up an important contribution to the human diet in many countries because fermentation is an inexpensive technology, which preserves the food, improves its nutritional value and enhances its sensory properties (Gadaga, Mutukumira, Narvhus, & Feresu, 1999). Traditional fermented beverages have been documented in di?erent countries, especially in Africa. Kule naoto, or Kwerionik, for example, are fermented milks produced and consumed in Kenya and Uganda, respectively (Pinto, Franz, Schillinger, & Holzapfel, 2006). Amasi (mukaka wakakora or zifa) is a traditional fermented milk consumed in Zimbabwe. It is produced by leaving fresh raw bovine milk to ferment naturally at ambient temperature in earthenware pots, or any other suitable containers. The microorganisms inherent in the milk, the container and the surrounding air are assumed to ferment the milk within 1–3 days, depending on the ambient temperature. From these, bacterial strains belonging to Lactococcus, Lactobacillus, Leuconostoc and Enterococcus have been identi?ed (Gadaga et al., 1999). Ergo is traditional fermented milk produced in Ethiopia. It is thick, smooth, of uniform appearance and has some resemblance to yoghurt. Usually it has a white milk color when prepared carefully. The product is semi-solid and has a pleasant odor and taste. Several microorganisms are present in ergo such as Gram-positive bacteria. Yeasts and moulds were also isolated but were not identi?ed. The LAB included Streptococcus thermophilus, Streptococcus acidominus, Streptococcus bovis, Streptococcus mitis, Streptococcus agalactiae, Enterococcus faecalis var. liquefaciens, Lactococcus cremoris, Lactococcus lactis, Leuconostoc dextranicum, Leuconostoc lactis and Lactobacillus xylosus
Fig. 2. Graphical representation of a nutraceutic.
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(Gonfa, Fite, Urga, & Gashe, 1999; Gonfa, Foster, & Holzapfel, 2001). The original observation of the positive role played by some selected bacteria is attributed to Eli Metchniko?, the Russian born Nobel Prize winner working at the Pasteur Institute at the beginning of the last century, who suggested that ‘‘the dependence of the intestinal microbes on the food makes it possible to adopt measures to modify the ?ora in human bodies and to replace the harmful microbes by useful microbes’’ (FAO/WHO, 2001). Almost at the same time, the ?rst formulae for deliberate administration of live lactic acid bacteria was launched in Paris, which was a yogurt based on a culture of Sc. thermophilus and Lactobacillus delbrueckii subsp. bulgaricus. With this successful introduction, the incorporation of probiotic bacteria in foods was focused on milk-based products (Molin, 2001). Macedo (1997) developed a probiotic beverage with a mix of 35% bu?alo cheese whey, 30% soymilk and 35% cow milk fermented by a mixed culture of Lactobacillus casei Shirota and Bi?dobacterium adolescentis. The concomitant use of these two strains resulted both in a pleasant taste and ?avoured product besides an expected association with health-promoting e?ects. The fermentation was carried out at 37 °C for 8 h keeping a 1:5 proportion between the lactic acid and bi?dobacteria in a 5% inoculum amount. The fermented beverage with a light extra-?avoring with vanilla essence was evaluated during a 28 days storage period at 4 °C, thus con?rming the preservation of its chemical, microbiological and sensorial stabilities, although, a slight variation in the acidity pro?le the desirable balance for the viable cells of the bacterial strains (6.8 · 108 cells/mL for Lb. casei Shirota and 2.3 · 107 cells/ mL for Bf. adolescentis) was found in the sensorial characteristics of the fermented milk beverage allowing its good acceptability in all time course of the storage period. Vinderola, Gueimonde, Delgado, Reinheimer, and De ? Los Reyes-Gavilan (2000) evaluated the carbonation of pasteurized milk as a method for improving the fermentation conditions and/or bacterial viability in the fermented milk added with Lactobacillus acidophilus and/or Bi?dobacterium bi?dum. The fermentation time was signi?cantly lowered in carbonated milk. The use of CO2 had no detrimental e?ects on the sensory properties of the fermented milk and did not exert any in?uence on the viability of Lb. acidophilus. Oliveira et al. (2002) studied the combined e?ect of milk supplementation and culture composition on acidi?cation, textural properties, and the microbiological stability of the fermented milks containing probiotic bacteria. Whey, casein hydrolysate and milk proteins were tested as supplementation. Two strains of probiotic bacteria, Lb. acidophilus LA5 and Lactobacillus rhamnosus LC35, were used in pure culture, and in mixed culture with Sc. thermophilus (ST7). The results showed that fermented products texture was not dependent on the culture composition, but strongly dependent on milk supplementation. Sodini, Lucas, Tissier, and Corrieu (2005) studied the physical
properties and the microstructure of the yoghurts containing probiotic bacteria and supplemented with milk protein hydrolysates. Three casein hydrolysates and three whey protein hydrolysates were added to the milk. The milks were fermented with either of the two di?erent cultures and compared to the control yoghurt without supplementation. For both the cultures, the addition of the hydrolysates reduced the fermentation time but negatively a?ected the texture, decreasing the complex viscosity and graininess in the yoghurts. The ingestion of the selected strains of the probiotic bacteria and the nutraceuticals has the protential to bene?t the consumer but little information is available regarding the e?ect of nutraceuticals on the viability of probiotic species of bacteria in a foodstu?. Awaisheh, Haddadin, and Robinson (2005) studied the e?ect of di?erent concentrations of three nutraceuticals (x-3-fatty acids, iso?avones and phytosterols) on the viability of two probiotic cultures (Lactobacillus gasseri and Bi?dobacterium infantis), which were incorporated into the yoghurt. The sensory qualities of the yoghurt and the viability of the probiotic species were evaluate during the storage at 4 °C. The combination of the three nutraceuticals gave signi?cantly higher counts than the control milk. In particular, Bf. infantis count was higher in the combination than in the presence of individual nutraceuticals. Some strains of LAB as Lactobacillus ssp. SLH6 and Sc. thermophilus ST4 were used as unocula to ferment experimental mixtures of milk powder (3 g%) plus cassava starch (6 g%), or inulin (6 g%) as prebiotic. The use of this type of milk–oligosaccharides mixtures o?ers the possibility to reduce the use of milk to 1/3 of that used in regular yogurts, which can also serve as vehicles for the probiotic and prebiotic agents. The product is yogurt-like and the taste is fairly good. It can be o?ered to those countries that have shortage of milk and are good producers of cassava starch and other starchy products. The bioavailability of many nutrients in vegetable diets is usually low, and this will signi?cantly contribute to the nutritional inadequacy. In poor countries animal foods and milk although of excellent nutritional value, are not available in enough quantity to these populations, due to their customs and high coasts (Parada, Sambucetti, Zuleta, & Rio, 2003). The e?ect of ra?nose family oligosaccharides (RFOs) addition on probiotic viability of Bi?dobacterium lactis Bb-12 and Lb. acidophilus La-5 in the fermented milk ? ? ? was evaluated by Mart?nez-Villaluenga, Fr?as, Gomez, and Vidal-Valverde (2006) during 21 days of storage at 5 °C. Viability retention of both strains were higher in the fermented milk with RFOs, which had bene?cial e?ects on the survival of these probiotic cultures in the dairy products. Stored products containing the probiotics and the prebiotics have synergistic actions in the promotion of health. Evolus? (Valio, Finland) is the ?rst blood pressure reducing fermented milk drink developed in Finland in October 2000. Its bene?cial e?ect is due to the bioactive
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peptides that are generated when Lactobacillus helveticus decomposes milk casein. A host of clinical tests have shown these peptides to have a positive in?uence on lowering the blood pressure (Valio, 2006). Faced with the market saturation in the third world and stock dumping in advanced countries, dairy organizations that produce traditional fermented products must explore new avenues of the product diversi?cation. New trends are becoming apparent with regard to consumers acceptance of the fermented milks. The selection of the microorganisms used as starters in the manufacture of the fermented milks is now based on the interaction with the gastrointestinal micro?ora and the metabolic properties. Genetic engineering of the microorganisms and new technological possibilities is expected to create new fermented milks in which desired properties would be emphasized. New products should be developed and traditional ones should be maintained and improved. Hence, the introduction of new technologies assumes added signi?cance. Other potential areas include special fermentation products, such as probiotic milk foods with immunomodulating activity. These products could receive greater consumer attention as the market demand for the value-added fermented products is expected to be very high in the near future. Similarly, Bi?dobacterium-derived fermented products have shown excellent physiological activity in infant digestion metabolism and nutrient utilization. Milk-based formulae with bioactive peptides of immunological signi?cance can be prepared by fermentation process to mimic the biological attributes of breast milk. These examples are only few applications of the potentiality of probiotics. However, other vehicles such as nondairy raw materials may be used to deliver into the host the bene?c lactic acid bacteria and other microorganism. Evidently, the fermented milk products continue to be subject of intensive researches to enhance their inherent nutritional and healthful attributes. Unsoundly, these products represent an exciting area of commercial interest for the dairy industry. The commercial stability of the fermented milks allows the expansion and coverage of the needs of di?erent consumer categories. With the growing interest of consumers in health-related foods, the market for probiotic health microorganisms would have a bright and promising future (Batish & Grover, 2004). 4. Non-dairy probiotic beverages Probiotic products are usually marketed in the form of fermented milks and yoghurts; however, with an increase in the consumer vegetarianism throughout the developed countries, there is also a demand for the vegetarian probiotic products. Furthermore, lactose intolerance and the cholesterol content are two major drawbacks related to the fermented dairy products (Heenan, Adams, Hosken, & Fleet, 2004; Yoon, Woodams, & Hang, 2006). There are a wide variety of traditional non-dairy fermented beverages produced around the world. Much of them are
non-alcoholic beverages manufactured with cereals as principal raw material. Boza is a beverage consumed in Bulgaria, Albania, Turkey and Romania. It is a colloid suspension, from light to dark beige, sweet, slightly sharp to slightly sour, made from wheat, rye, millet, maize and other cereals mixed with sugar, or saccharine. Boza has become a very popular beverage consumed daily by people of all ages due to its pleasant taste, ?avour and high nutritional values. Micro?ora identi?cation of Bulgarian boza shows that it mainly consists of yeasts and lactic acid bacteria, in an average LAB/yeast ratio of 2.4. The lactic acid bacteria isolated were Lactobacillus plantarum, Lb. acidophilus, Lactobacillus fermentum, Lactobacillus coprophilus, Leuconostoc re?nolactis, Leuconostoc mesenteroides and Lactobacillus brevis. The yeasts isolated were Saccharomyces cerevisiae, Candida tropicalis, Candida glabrata, Geotrichum penicillatum and Geotrichum candidum (Blandino, Al-Aseeri, Pandiella, Cantero, & Webb, 2003; Gotcheva, Pandiella, Angelov, Roshkova, & Webb, 2000). Bushera is the most common traditional beverage prepared in the Western highlands of Uganda. The product is consumed by both the young children and the adults. The sorghum, or millet ?our from the germinated sorghum and millet grains is mixed with the boiling water and left to cool to ambient temperature. Germinated millet or sorghum ?our is then added and the mixture is left to ferment at ambient temperature for 1–6 days. The lactic acid bacteria isolated from Bushera comprised of ?ve genera, Lactobacillus, Lactococcus, Leuconostoc, Enterococcus and Streptococcus. Lb. brevis was more frequently isolated than other species (Muianja, Narvhus, Treimo, & Langsrud, 2003). Mahewu (amahewu) is a sour beverage made from the corn meal, consumed in Africa and some Arabian Gulf countries. It is prepared from the maize porridge, which is mixed with the water. The sorghum, millet malt, or wheat ?our is then added and left to ferment. The spontaneous fermentation process is carried out by the natural ?ora of the malt at the ambient temperature. The predominant microorganism found in African mahewu is Lactococcus lactis subsp. lactis (Blandino et al., 2003; Gadaga et al., 1999). Pozol is a refreshing beverage, consumed in the Southeastern Mexico, made by cooking maize in an approximately 1% (w/v) lime solution, washing with water, grinding to make a dough known as nixtamal, shaping into balls, wrapping in banana leaves and leaving to ferment at ambient temperature for 0.5–4 days. The fermented dough is suspended in he water and drunk. Some ?brous components are not completely solubilized by nixtamalization and sediment is present in the beverage when the dough is suspended in the water (Wacher et al., 2000). Togwa is a starch-sacchari?ed traditional beverage consumed in Africa. In the southern part of Tanzania, togwa is usually made from the maize ?our and ?nger millet malt. In this region, it is consumed by the working people and
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also used as a refreshment as well as a weaning food (Oi & Kitabatake, 2003). Togwa is also regularly consumed by the young children. The cereal, or cassava ?our is cooked in the water. After cooling at 35 °C, starter culture (old togwa) and cereal ?our from the germinated grains are added. The fermentation process ?nishes at pH 4.0–3.2 (Molin, 2001). Lactic acid bacteria can rarely convert starch into lactic acid. However, some strains of Lactobacillus and Streptococcus can do it (Parada, Zapata, De Frabrizio, & Martinez, 1996). For example, Lb. plantarum A6, isolated by Giraud, Grosselin, Parada, and Raimbault (1993), showed extracellular amylase activity. Sour cassava starch is obtained by a natural fermentation and this product is largely appreciated in Africa, South America and other developing countries. Lb. plantarum D34 was isolated in Colombia, which showed an amylolytic property. Santos (2001) developed a probiotic beverage with the fermented cassava ?our (liquid phase + solid) using mixed culture of Lb. plantarum A6, which were amylolytic strains of Lb. casei Shirota and Lb. acidophilus. The best parameters of the fermentation were 8% inoculation rate, incubation temperature and period as 35 °C and 16 h, respectively, and 20% of cassava ?our. At the end of the fermentation, the amount of bacteria was 2.8 · 109 cells/mL of lactic amylolytic bacteria and 2.3 · 109 cells/mL of probiotic bacteria. The ?nal lactic beverage had 36% of guava juice, 10% of sugar and 54% of fermented lactic beverage. The lactic beverage maintained its microbiological and physico-chemical quality for 28 days storage period at 4 °C. Angelov, Gotcheva, Kuncheva, and Hristozova (2006) produced a symbiotic functional drink from the oats by combining a probiotic starter culture and whole-grain oat substrate. The oats and barley are the cereals with highest content of beta-glucan, recognized as the main functional component of the cereal ?bers. Studies have indicated the hypocholesterolemic e?ect of this compound, leading to 20–30% reduction of LDL-cholesterol, and to an expected overall e?ect of reduced cardiovascular disease risk (Stark & Madar, 1994; Wrick, 1994). The substrate was fermented with Lb. plantarum B28. The levels of starter culture concentration, oat ?our and sucrose content were established for completing a controlled fermentation for 8 h. The addition of aspartame, sodium cyclamate, saccharine and Huxol (12% cyclamate and 1.2% saccharine) had no e?ect on the dynamics of the fermentation process and on the viability of the starter culture during the product storage. The beta-glucan content in the drink of 0.31–0.36% remained unchanged throughout fermentation and storage of the drink. The viable cells counts reached at the end of the process were about 7.5 · 1010 CFU/mL. The shelf life of the oat drink was estimated to 21 days under refrigerated storage. Since few decades ago, the soybean has received attention from the researchers due to its protein quality. Because of their functional properties, it has a great application potential in the food industry. The soymilk is suitable for
the growth of the lactic acid bacteria, especially bi?dobacteria (Chou & Hou, 2000; Matsuyama et al., 1992). Several studies have mentioned the production and use of the fermented soymilk drinks as probiotic, mainly soybean yogurt, which further can be supplemented with oligofructose and inulin (Fuchs, Borsato, Bona, & Hauly, 2005; Shimakawa, Matsubara, Yuki, Ikeda, & Ishikawa, 2003; Wang, Yu, & Chou, 2002). Despite potential sensory challenges, there is a genuine interest in the development of fruit-juice based functional beverages, forti?ed with the probiotic and prebiotic ingredients. The fruit juices have been suggested as an ideal medium for the functional health ingredients because they inherently contain bene?cial nutrients, they have taste pro?les that are pleasing to all the age groups, and because they are perceived as being healthy and refreshing (Tuorila & Cardello, 2002). The fruits and vegetables are rich in the functional food components such as minerals, vitamins, dietary ?bers, antioxidants, and do not contain any dairy allergens that might prevent usage by certain segments of the population (Luckow & Delahunty, 2004a). Hardaliye is a lactic acid fermented beverage produced from the natural fermentation of the red grape, or grape juice with the addition of the crushed mustard seeds and benzoic acid. This beverage can be found in the Thrace region of Turkey. It is very well known and has been produced and consumed since ancient times. The mustard seed’s eteric oils a?ect the yeasts and also give ?avor to the ?nal product. Benzoic acid inhibits, or decreases alcohol production by a?ecting the yeast. Once fermented, the hardaliye is stored at a temperature of 4 °C and consumed either fresh, or aged. The lactic acid bacteria found in the beverage were Lactobacillus paracasei subsp. paracasei, Lactobacillus casei subsp. pseudoplantarum, Lactobacillus brevis, Lactobacillus pontis, Lactobacillus acetotolerans, Lactobacillus sanfransisco and Lactobacillus vaccinostercus. This characterization allowed the selection of appropriate strains for the manufacture of hardaliye using pasteurized, or sterile ?ltered grape juices (Arici & Coskun, 2001). Yoon, Woodams, and Hang (2004) determined the suitability of the tomato juice as a raw material for the production of probiotic juice by Lb. acidophilus LA39, Lb. plantarum C3, L. casei A4 and Lb. delbrueckii D7. The tomato juice was inoculated with a 24 h-old culture and incubated at 30 °C. The lactic acid cultures reduced the pH to 4.1 and the viable cell counts reached nearly (1.0– 9.0) · 109 CFU/mL after 72 h fermentation. The viable cell counts of the four lactic acid bacteria in the fermented tomato juice raged from 106 to 108 CFU/mL after 4 weeks of cold storage at 4 °C. Yoon, Woodams, and Hang (2005) also evaluated the potential of red beets as the substrate for the production of probiotic beet juice by the above four species of lactic acid bacteria. All the lactic cultures were capable of rapidly utilizing the beet juice for the cell synthesis and lactic acid production. Lb. acidophilus and Lb. plantarum produced higher amount of lactic acid than other cultures and reduced the pH of the fermented beet juice from
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an initial value of 6.3 to below 4.5 after 48 h of fermentation at 30 °C. Although the lactic cultures in fermented beet juice gradually lost their viability during the cold storage, the viable cell counts of these bacteria, except for Lb. acidophilus, in the fermented beet juice still remained at 106–108 CFU/ mL after 4 weeks of cold storage at 4 °C. Yoon et al. (2006) also developed a probiotic cabbage juice using lactic acid bacteria. Cabbage juice was inoculated with a 24 h-old lactic culture and incubated at 30 °C. The cultures (Lb. plantarum C3, Lb. casei A4 and Lb. delbrueckii D7) grew well on cabbage juice and reached about 1 · 109 CFU/mL after 48 h of the fermentation. Lb. casei produced a lower amount of titratable acidity, expressed as lactic acid, than Lb. delbrueckii, or Lb. plantarum. After 4 weeks of the cold storage at 4 °C, the viable cell counts of Lb. plantarum and Lb. delbrueckii were 4.1 · 107 and 4.5 · 105 CFU/mL, respectively. Lb. casei did not survive at low pH and lost cell viability completely after 2 weeks of the cold storage. The fermented cabbage juice could serve as a healthy beverage for vegetarians and lactose-allergic consumers. Rakin, Vukasinovic, Siler-Marinkovic, and Maksimovic (2007) enriched beetroot and carrot juices with the brewer’s yeast autolysate before lactic acid fermentation with Lb. acidophilus. The addition of the autolysate favorably a?ected the increase of the number of lactic acid bacteria during the fermentation (Aeschlimann & Von Stocar, 1990), reduction of the time of fermentation and enrichment of the vegetable juices with amino acids, vitamins, minerals and antioxidants (Chae, Joo, & In, 2001). The use of spent brewer’s yeast from the brewery was important for the economic optimization of the fermentation. A mixture of beetroot and carrot juices has optimum proportions of pigments, vitamins and minerals (Rakin et al., 2007). Luckow and Delahunty (2004a) evaluated the consumer’s acceptance for the appearance, aroma, texture and ?avour of the probiotic fruit juices. Novel blackcurrant juices containing the probiotic cultures were compared with the conventional blackcurrant juices by the descriptive analysis. The probiotic juices contained aroma and ?avors characteristic of the functional ingredients. Subsequent testing took place in a local shopping center, where the consumers were presented with two randomly coded blackcurrant juice samples. One of the products was a natural blackcurrant juice, and the other was a commercially processed blackcurrant juice containing probiotic cultures. The consumers were instructed that one of the juice samples contained ‘‘special ingredients’’ designed to improve their health. They were asked to assess their overall impression of both the juices, and to rate their acceptance of the sensory characteristics. Furthermore, based on their overall impressions and guided by their individual expectations, the consumers were asked to identify the juice they perceived to be the ‘healthiest’ (e.g., containing the ‘‘special ingredients’’). The juice preference was dependent on the gender and the age. In general, the consumers selected their most preferred juice product as the ‘healthiest’ sample.
Luckow and Delahunty (2004b) examined the sensory impact of functional ingredients (e.g., probiotics, prebiotics, vitamins, and minerals) on the aroma and taste of orange fruit juices. A trained panel (n = 10) performed a descriptive sensory analysis on four functional orange juices and seven conventional orange juices. The functional juices were described as possessing perceptible ‘‘dairy’’, ‘‘medicinal’’ and ‘‘dirty’’ ?avors, distinguishing them from the conventional juices. Subsequently, 100 consumers participated in a preference test, whereby ?ve orange juices (three conventional and two functional) were ranked in order of the consumer preference. The ranking decisions were based solely on the sensory characteristics of the juices, since product information was not provided. On an overall basis, the consumers preferred the sensory characteristics of conventional juices. However, cluster analysis identi?ed a small consumer segment (11%) that signi?cantly preferred the sensory attributes of functional juices. Evidently, there are already some relatively new nondairy probiotic beverages in the market. Grain?elds Wholegrain Liquid? is a refreshing, e?ervescent liquid that delivers active, friendly lactic acid bacteria and yeasts as well as vitamins, amino acids, and enzymes. It is made from organic ingredients including the grains, beans, and seeds such as the malted organic oats, maize, rice, alfalfa seed, pearl barley, linseed, mung beans, rye grain, wheat, millet. The liquid is fermented to achieve high levels of active probiotic bacteria sustained in a liquid medium immediately available for the use within the digestive system. Grain?elds Wholegrain Liquid? is fermented with lactobacili and yeasts cultures: Lb. acidophilus, Lb. delbreukii, Saccharomyces boulardii and Sc. cerevisiae. The liquid is dairy-free, contains no genetically modi?ed ingredients and has no added sugar (Superfoods., 2006). Vita Biosa? is a mixture of aromatic herbs and other plants, which are fermented by a combination of lactic acid and yeast cultures. This beverage contains no sugar and can produce carbon dioxide. The bacteria are selected based on the criterion of their providing the micro?ora of the intestines with the best possible conditions. During the fermentation, the lactic acid formed produces a low pH value of about 3.5. The low pH prevents the development of harmful bacteria in the ?nished product. Vita Biosa? also contains a high number of antioxidants. The reason for this beverage becoming so popular is its ability to quickly restore the natural balance in the digestive system. The liquid restricts harmful bacteria and thus gives the bene?cial bacteria a better possibility to multiply and create healthy intestinal ?ora. Vita Biosa? is manufactured in Denmark (Superfoods, 2006). Proviva? was the ?rst probiotic food that does not contain milk, or milk constituents. It was launched in Sweden by Skane Dairy (Sweden) in 1994. The active component comprises lactic acid bacteria fermented oatmeal gruel. Malted barely is added to enhance the liquefaction of the product and Lb. plantarum 299v carries out the fermentation. The ?nal product contains 1 · 1012 colony-forming
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units (CFU) of L. plantarum/L. This formula is used as the active ingredient in the food product in which 5% of the oat meal gruel is mixed with a fruit drink. The consumer product contains 5 · 1010 CFU/L (Molin, 2001). Valio Ltd. began developing non-dairy drinks with Lb. rhamnosus GG in 1996 and the ?rst product was launched in 1997. Ge?lus? fruit drinks have a shelf life of 5 weeks when refrigerated (Leporanta, 2005a, 2005b). The latter was from Valio’s commercial Biopro?t? product, with Lb. rhamnosus GG and Propionibacterium freudenreichii ssp. shermanii JS (Daniells, 2006). Biola? juice drink is a new probiotic juice manufactured by Tine BA in Norway. It contains added Lb. rhamnosus GG. Tine is using this strain under license from Finnish dairy company Valio Ltd. The juice drinks contain more than 95% fruit and no added sugar and are available in orange–mango and apple–pear ?avours (Leporanta, 2005b). Rela? is a fruit juice with Lactobacillus reuteri MM53 manufactured by the Biogaia, Sweden. In recent years, the consumer demand for non-dairybased probiotic products has increased (Shah, 2001) and the application of probiotic cultures in non-dairy products and environments represents a great challenge (MattilaSandholm et al., 2002). Juice manufacturers in particular were considered to lead new product development activities for gut bene?t beverages as line extensions of existing functional drinks such as Valio’s Ge?lus?. Fruit juice-based probiotic drinks would become an increasingly important category in future years (Dairy Industries International, 2004; Leatherhead Food Research Association, 2004). 5. Perspectives of probiotic beverages There is every reason to believe that beverages would be the next food category where the healthy bacteria will make their mark. Likely candidates are chilled fruit juices, bottled water, or fermented vegetable juices. The probiotic microorganisms also have been directly incorporated into the drinks. The key to the development of this second generation of the probiotic products is a special Direct Liquid Inoculation system. It allows food producers to add the probiotic bacteria directly to the ?nished food product. The technology uses Tetra Pak’s aseptic dosing machine Flex Dos that allows the bacteria to be added to liquids just before they are ?lled into the cartons. The innovation is expected to signi?cantly boost the market for the probiotic beverages, which have so far been restricted by the delicate nature of the ingredient and concerns over the contamination. The probiotic bacteria must be live to exert their health bene?ts but they can be destroyed by a number of processing situations. In beverages, for example, the heat treatment in a standard production run would kill the live bacteria (Chr Hansen, 2006; Shah, 2001). Adding probiotics to the juices is more complex than formulating in the diary products because the bacteria need protection from the acidic conditions in the fruit juice. However, with microencapsulation technologies, the probi-
otics can become an important ingredient in the functional foods, expanding the probiotic application outside the pharmaceutical and supplement industries. Microencapsulation technologies have been developed and successfully applied using various matrices to protect the bacterial cells from the damage caused by the external environment (Del Piano et al., 2006). It is the process by which small particles or droplets are surrounded by a coating to produce the capsules in the micrometer to millimeter range known as microcapsules. The microencapsulation allows the probiotic bacteria to be separated from its environment by a protective coating. Several studies have reported the technique of the microencapsulation by using gelatin, or vegetable gum to provide protection to acid-sensitive Bi?dobacterium and Lactobacillus (Chandramouli, Kailasapathy, Peiris, & Jones, 2004; Lee, Cha, & Park, 2004; O’Riordan, Andrews, Buckle and Conway, 2001; Sultana et al., 2000). New product development requires detailed knowledge of both the products and the customers. The high reported failure rates for new international functional beverages suggest a failure to manage the customer knowledge e?ectively, as well as a lack of the knowledge management between the functional disciplines involved in the new product development process. The methodologies that advance both a ?rm’s understanding of the customer’s choice motives and the value systems, and its knowledge management process, can increase the chances of new product success in the international functional beverages market (Bogue & Sorenson, 2006). The commercial success of the probiotic products ultimately depends on taste and appeal to the consumer (Heenan et al., 2004; Yoon et al., 2006). The consumer needs to receive a comprehensible and reasonable message about probiotics, without appearing to be exaggerated. The ‘‘health claims’’ must be defensible when placed under the scrutiny by the controlling authorities. In the coming years of the new Millennium, changes would occur in the interface between the scienti?c studies and the acceptance by the consumers (Gorbach, 2002). With increasingly competitive markets, the functional foods and the beverages manufacturers have targeted functionality as an extremely important marketing tool to create competitive advantages in the marketplace (Sorenson & Bogue, 2005). References
Aeschlimann, A., & Von Stocar, U. (1990). The e?ect of yeast extract supplementation on the production of lactic acid from whey permeate by Lactobacillus helveticus. Applied Microbiology and Biotechnology, 32, 398–440. Aguirre, M., & Collins, M. D. (1993). Lactic acid bacteria and human clinical infection. Journal of Applied Bacteriology, 75, 95–107. Aiba, Y., Suzuki, N., Kabir, A. M. A., Takagi, A., & Koga, Y. (1998). Lactic acid-mediated suppression of Helicobacter pylori by the oral administration of Lactobacillus salivarius as probiotic in a gnotobiotic murine model. American Journal of Gastroenterology, 93, 2097–2101. Angelov, A., Gotcheva, V., Kuncheva, R., & Hristozova, T. (2006). Development of a new oat-based probiotic drink. International Journal of Food Microbiology, 112, 75–80.
F.C. Prado et al. / Food Research International 41 (2008) 111–123 Arici, M., & Coskun, F. (2001). Hardaliye: Fermented grape juice as a traditional Turkish beverage. Food Microbiology, 18, 417–421. Arunachalam, K., Gill, H. S., & Chandra, R. K. (2000). Enhancement of natural immunity function by dietary consumption of Bi?dobacterium lactis HN019. European Journal of Clinical Nutrition, 54, 1–4. Aso, Y., Akazan, H., Kotake, T., Tsukamoto, T., Imai, K., & Naito, S. (1995). Preventive e?ect of a Lactobacillus casei preparation on the recurrence of super?cial bladder cancer in a double-blind trial. European Urology, 27, 104–109. Awaisheh, S. S., Haddadin, M. S. Y., & Robinson, R. K. (2005). Incorporation of selected nutraceuticals and probiotic bacteria into a fermented milk. International Dairy Journal, 15, 1184–1190. Batish, V. K., & Grover, S. (2004). Fermented milk products. In A. Pandey (Ed.), Concise encyclopedia of bioresource technology (pp. 201–209). USA: The Haworth Press. Blandino, A., Al-Aseeri, M. E., Pandiella, S. S., Cantero, D., & Webb, C. (2003). Cereal-based fermented foods and beverages. Food Research International, 36, 527–543. Bogue, J., & Sorenson, D. (2006). In Proceedings of the 98th EAAE (European Association of the Agricultural Economists) Seminar. Product optimization in new product development: The case of innovative probiotic beverages. ? Brasil. (1999a). Portaria no. 15, de 30 de abril de 1999. Diario O?cial da ? Uniao, Bras?lia, 14 maio 1999. ? ? Brasil. (1999b). Resolucao no. 16, de 30 de abril de 1999. Diario O?cial da ?? ? Uniao, Bras?lia, 03 dez. 1999. ? ? Brasil. (1999c). Resolucao no. 18, de 30 de abril de 1999. Diario O?cial da ?? ? Uniao, Bras?lia, 03 maio 1999. ? ? Brasil. (1999d). Portaria no. 19, de 30 de abril de 1999. Diario O?cial da ? Uniao, Bras?lia, 03 maio 1999. ? Chae, H. J., Joo, H., & In, M. J. (2001). Utilization of brewer’s yeast cells for the production of food grade yeast extract. Part I: E?ects of di?erent enzymatic treatments on solid and protein recovery and ?avor characteristics. Bioresource Technology, 76, 253–258. Chandramouli, V., Kailasapathy, K., Peiris, P., & Jones, M. (2004). An improved method of microencapsulation and its evaluation to protect Lactobacillus spp. in simulated gastric conditions. Journal of Microbiological Methods, 56, 27–35. Chou, C. C., & Hou, J. W. (2000). Growth of bi?dobacteria in soymilk and survival in the fermented soymilk drink during storage. International Journal of Food Microbiology, 56, 113–121. Chr Hansen. (2006). Probiotics – Products – Beverages. Available at www.chr-hansen.com (accessed 20 June 2006). ? Coconnier, M. H., Lievin, V., Hemery, E., & Servin, A. L. (1998). Antagonistic activity against Helicobacter infection in vitro and in vivo by the human Lactobacillus acidophilus strain LB. Applied and Environmental Microbiology, 64, 4573–4580. Collins, J. K., Thornton, G., & O’Sullivan, G. O. (1998). Selection of probiotic strains for human applications. International Dairy Journal, 8, 487–490. Crittenden, R. G. (1999). Prebiotics. In G. W. Tannock (Ed.), Probiotics: A critical review (pp. 141–156). Wymondham, UK: Horizon Scienti?c Press. Dairy Industries International. (2004). Drinking to health (Vol. 69, p. 14). Daniells, S. (2006). Valio continues research into probiotic fruit juices (14/ 04/2006). Nutraingredients. Available at www.nutraingredients.com (accessed in 28 February 2007). ? De Bondt, V. (2003). Novas tendencias para bebidas funcionais. Brasil Alimentos, 18, 26–27. De Roos, N. M., & Katan, M. B. (2000). E?ects of probiotic bacteria on diarrhoea, lipid metabolism, and carcinogenesis: a review of papers published between 1988 and 1998. American Journal of Clinical Nutrition, 71, 405–411. Del Piano, M., Morelli, L., Strozzi, G. P., Allesina, S., Barba, M., Deidda, F., et al. (2006). Probiotics: From research to consumer. Digestive and Liver Diseases, 38(Suppl. 2), S248–S255. Donnet-Hughes, A., Rochat, F., Serrant, P., Aeschlimann, J. M., & Schi?rin, E. J. (1999). Modulation of nonspeci?c mechanisms of
defense by lactic acid bacteria: E?ective dose. Journal of Dairy Science, 82, 863–869. El-Nezami, H., Mykkanen, H., Kankaanpaa, P., Salminen, S., & Ahokas, ¨ ¨¨ J. (2000). Ability of Lactobacillus and Probionibacterium strains to remove a?atoxin B1 from chicken duodenum. Journal of Food Protect, 63, 549–552. FAO/WHO. (2001). Health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria. Cordoba, Argentina: Food and Agriculture Organization of the United Nations and World Health Organization Expert Consultation Report. Finley, J. W. (1996). Designer foods. Is there a role for supplementation/ forti?cation? Dietary phytochemicals in cancer prevention and treatment. New York, EUA: American Institute for Cancer Research. Plenum Press (Chapter 9). Fuchs, R. H. B., Borsato, D., Bona, E., & Hauly, M. C. O. (2005). ? ‘‘Iogurte’’ de soja suplementado com oligofrutose e inulina. Ciencia e Tecnologia de Alimentos, 25, 175–181. Gadaga, T. H., Mutukumira, A. N., Narvhus, J. A., & Feresu, S. B. (1999). A review of traditional fermented foods and beverages of Zimbabwe. International Journal of Food Microbiology, 53, 1–11. Gibson, G. R., & Roberfroid, M. B. (1995). Dietary modulation of the human colonic microbiota: Introducing the concept of prebiotics. Journal of Nutrition, 125, 1401–1412. Gill, H. S., Cross, M. L., Rutherfurd, K. J., & Gopal, P. K. (2001). Dietary probiotic supplementation to enhance cellular immunity in the elderly. British Journal of Biomedical Science, 57, 94–96. Gill, H. S., Rutherfurd, K. J., Prasad, J., & Gopal, P. K. (2000). Enhancement of natural and acquired immunity by Lactobacillus rhamnosus (HN001), Lactobacillus acidophilus (HN017) and Bi?dobacterium lactis (HN019). British Journal of Nutrition, 83, 167–176. Giochetti, P., Rizzello, F., Venturi, A., Brigidi, P., Matteuzzi, D., Bazzocchi, G., et al. (2000). Oral bacteriotherapy as maintenance treatment in patients with chronic pouchitis: A double-blind, placebocontrolled trial. Gastroenterology, 119, 305–309. Giraud, E., Grosselin, L., Parada, J. L., & Raimbault, M. (1993). Puri?cation and characterization of an extracellular amylase from Lactobacillus plantarum strain A6. Journal of Applied Bacteriology, 75, 276–282. Gonfa, A., Fite, A., Urga, K., & Gashe, B. A. (1999). The microbiological aspects of ergo (ititu) fermentation. Ethiopian Journal of Science, 22, 2. Gonfa, A., Foster, H. A., & Holzapfel, W. H. (2001). Field survey and literature review on traditional fermented milk products of Ethiopia. International Journal of Food Microbiology, 68, 173–186. Gorbach, S. L. (2002). Probiotics in the third millennium. Digestive and Liver Diseases, 34(Suppl. 2), S2–S7. Gotcheva, V., Pandiella, S. S., Angelov, A., Roshkova, Z. G., & Webb, C. (2000). Micro?ora identi?cation of the Bulgarian cereal-based fermented beverage boza. Process Biochemistry, 36, 127–130. Guandalini, S., Pensabene, L., Zikri, M. A., Dias, J. A., Casali, L. G., Hoekstra, H., et al. (2000). Lactobacillus GG administered in oral rehydration solution to children with acute diarrhoea: A multicenter European trial. Journal of Pediatric and Gastroenterology Nutrition, 30, 54–60. Guarino, A., Berni, C. R., Spagnuolo, M. I., Albano, F., & Di Benedetto, L. (1997). Oral bacterial therapy reduces the duration of symptoms and of viral excretion in children with mild diarrhoea. Journal of Pediatric and Gastroenterology Nutrition, 25, 516–519. Guarner, F., & Schaafsma, G. J. (1998). Probiotics. International Journal of Food Microbiology, 39, 237–238. Gupta, P., Andrew, H., Kirschner, B. S., & Guandalini, S. (2000). Is Lactobacillus GG helpful in children with Crohn’s disease? Results of a preliminary, open-label study. Journal of Pediatric and Gastroenterology Nutrition, 31, 453–457. Havenaar, R., & Huis In’t Veld, J. H. J. (1992). Probiotics: A general view. In B. J. B. Wood (Ed.). The lactic acid bacteria. 1 – The lactic acid bacteria in health and disease (pp. 209–224). New York, EUA: Chapman & Hall Press.
F.C. Prado et al. / Food Research International 41 (2008) 111–123 probiotic survival in fermented milk during refrigerated storage. International Dairy Journal, 16, 768–774. Matsuyama, J., Hirata, H., Yamagisi, T., Hayashi, K., Hirano, Y., Kuwata, K., et al. (1992). Fermentation pro?les and utilization of sugars of bi?dobacteria in soymilk. Nippon Shokuhin Kogyo Gakkaishi, 39, 887–893. Matsuzaki, T., & Chin, J. (2000). Modulating immune responses with probiotic bacteria. Immunology and Cell Biology, 78, 67–73. Mattila-Sandholm, T., Myllarinen, P., Crittenden, R., Mogensen, G., ¨ ? Fonden, R., & Saarela, M. (2002). Technological challenges for future probiotic foods. International Dairy Journal, 12, 173–182. Michetti, P., Dorta, G., Wiesel, P. H., Brassart, D., Verdu, E., Herranz, M., et al. (1999). E?ect of whey-based culture supernatant of Lactobacillus acidophilus (johnsonii) La1 on Helicobacter pylori infection in humans. Digestion, 60, 203–209. Midolo, P. D., Lambert, J. R., Hull, R., Luo, F., & Grayson, M. L. (1995). In vitro inhibition of Helicobacter pylori NCTC 11637 by organic acids and lactic acid bacteria. Journal of Applied Bacteriology, 79, 475–479. Molin, G. (2001). Probiotics in foods not containing milk or milk constituents, with special reference to Lactobacillus plantarum 299v. American Journal of Clinical Nutrition, 73(Suppl.), 380S–385S. Muianja, C. M. B. K., Narvhus, J. A., Treimo, J., & Langsrud, T. (2003). ? Isolation, characterisation and identi?cation of lactic acid bacteria from bushera: A Ugandan traditional fermented beverage. International Journal of Food Microbiology, 80, 201–210. Oatley, J. T., Rarick, M. D., Ji, G. E., & Linz, J. E. (2000). Binding of a?atoxin B1 to bi?dobacteria in vitro. Journal of Food Protection, 63, 1133–1136. Oi, Y., & Kitabatake, N. (2003). Chemical composition of an East African traditional beverage, togwa. Journal of Agricultural and Food Chemistry, 51, 7024–7028. Oliveira, M. N., Sivieri, K., Alegro, J. H. A., & Saad, S. M. I. (2002). ? ? Aspectos tecnologicos de alimentos funcionais contendo probioticos. ? ? Revista Brasileira de Ciencias Farmaceuticas, 38, 1–21. O’Riordan, K., Andrews, D., Buckle, K., & Conway, P. (2001). Evaluation of microencapsulation of a Bi?dobacterium strain with starch as an approach to prolonging viability during storage. Journal of Applied Microbiology, 91, 1059–1066. O’Sullivan, M. G., Thornton, G. M., O’Sullivan, G. C., & Collins, J. K. (1992). Probiotic bacteria: Myth or reality. Trends in Food Science and Technology, 3, 309–314. Oxman, T., Shapira, M., Klein, R., Avazov, N., & Rabinowitz, B. (2001). Oral administration of Lactobacillus induces cardioprotection. Journal of Alternative and Complementary Medicine, 7, 345–354. Parada, J. L., Sambucetti, M. E., Zuleta, A., & Rio, M. E. (2003). Lactic acid fermented products as vehicles for probiotics. In S. Roussos, C. R. Soccol, A. Pandey, & C. Augur (Eds.), New horizons in biotechnology (pp. 335–351). The Netherlands: Kluwer Academic Publishers. Parada, J. L., Zapata, L. E., De Frabrizio, S. V., & Martinez, A. (1996). Microbiological and technological aspects of cassava starch fermentation. World Journal of Microbiology and Biotechnology, 12, 53–56. Perdigon, G., Vintini, E., Alvarez, S., Medina, M., & Medici, M. (1999). Study of the possible mechanisms involved in the mucosal immune system activation by lactic acid Bacteria. Journal of Dairy Science, 82, 1108–1114. Perdone, C. A., Bernabeu, A. O., Postaire, E. R., Bouley, C. F., & Reinert, P. (1999). The e?ect of supplementation by Lactobacillus casei (strain DN-114 001) on acute diarrhoea in children attending day care centers. International Journal of Clinical Practices, 53, 179–184. Pinto, M. G. V., Franz, C. M. A. P., Schillinger, U., & Holzapfel, W. H. (2006). Lactobacillus ssp. with in vitro probiotic properties from human faeces and traditional fermented products. International Journal of Food Microbiology, 109, 205–214. Rakin, M., Vukasinovic, M., Siler-Marinkovic, S., & Maksimovic, M. (2007). Contribution of lactic acid fermentation to improved nutritive
Heenan, C. N., Adams, M. C., Hosken, R. W., & Fleet, G. H. (2004). Survival and sensory acceptability of probiotic microorganisms in a nonfermented frozen vegetarian dessert. Lebensmittel-Wissenschaft und-Technology, 37, 461–466. Hilliam, M. (2000). Functional food. How big is the market? The World of Food Ingredients, 12, 50–53. Hilton, E., Isenberg, H. D., Alperstein, P., France, K., & Borenstein, M. T. (1992). Ingestion of yogurt containing Lactobacillus acidophilus as prophylaxis for candidal vaginitis. Annals of International Medicine, 116, 353–357. Hilton, E., Rindos, P., & Isenberg, H. D. (1995). Lactobacillus GG vaginal suppositories and vaginitis. Journal of Clinical Microbiology, 33, 1433. Holzapfel, W. H., & Schillinger, U. (2002). Introduction to pre- and probiotics. Food Research International, 35, 109–116. Hosada, M., Hashimoto, H., He, D., Morita, H., & Hosono, A. (1996). E?ect of administration of milk fermented with Lactobacillus acidophilus LA-2 on faecal mutagenicity and micro?ora in human intestine. Journal of Dairy Science, 79, 745–749. Isolauri, E., Arvola, T., Sutas, Y., Moilanen, E., & Salminen, S. (2000). Probiotics in the management of atopic eczema. Clinical and Experimental Allergy, 30, 1604–1610. Isolauri, E., Juntunen, M., Rautanen, T., Sillanaukee, P., & Koivula, T. (1991). A human Lactobacillus strain (Lactobacillus casei sp. strain GG) promotes recovery from acute diarrhea in children. Pediatrics, 88, 90–97. Jack, R. V., Tagg, J. R., & Ray, B. (1995). Bacteriocins of gram-positive bacteria. Microbiology Reviews, 59, 171–200. Kabir, A. M., Aiba, Y., Takagi, A., Kamiya, S., Miwa, T., & Koga, Y. (1997). Prevention of Helicobacter pylori infection by lactobacilli in a gnotobiotic murine model. Gut, 41, 49–55. Kalliomaki, M., Salminen, S., Arvilommi, H., Kero, P., Koskinen, P., & Isolauri, E. (2001). Probiotics in primary prevention of atopic disease: A randomized placebo-controlled trial. Lancet, 357, 1076–1079. Leatherhead Food Research Association. (2004). Functional food markets, innovation and prospects: An international analysis (2nd ed.). Leatherhead International. Lee, J. S., Cha, D. S., & Park, H. J. (2004). Survival of freeze-dried Lactobacillus bulgaricus KFRI 673 in chitosan-coated calcium alginate microparticles. Journal of Agriculture and Food Chemistry, 52, 7300–7305. Leporanta, K. (2005a). Probiotics for juice-based products – Case Valio Ge?lus?. International sales, May 23, 2005. Available at www.valio.? (accessed in 05 January 2007). Leporanta, K. (2005b). Tine is using LGG under license from Valio Ltd. Valio Today’s – News, September 13, 2005. Available at www.valio.? (accessed in 05 January 2007). Luckow, T., & Delahunty, C. (2004a). Which juice is ‘healthier’? A consumer study of probiotic non-dairy juice drinks. Food Quality and Preference, 15, 751–759. Luckow, T., & Delahunty, C. (2004b). Consumer acceptance of orange juice containing functional ingredients. Food Research International, 37, 805–814. ? Macedo, R. E. F. (1997). Desenvolvimento de bebida lactea fermentada a ? ? base de extrato hidrossoluvel de soja e soro de leite de bufala por cultura mista de Lactobacillus casei Shirota e Bi?dobacterium adolescentis. MSc thesis. Brazil: UFPR, 141 pp. Majamaa, H., & Isolauri, E. (1996). Evaluation of the gut mucosal barrier: Evidence for increased antigen transfer in children with atopic eczema. Journal of Allergy and Clinical Immunology, 97, 985–990. Majamaa, H., & Isolauri, E. (1997). Probiotics: A novel approach in the management of food allergy. Journal of Allergy and Clinical Immunology, 99, 179–185. Majamaa, H., Isolauri, E., Saxelin, M., & Vesikari, T. (1995). Lactic acid bacteria in the treatment of acute rotavirus gastroenteritis. Journal of Pediatric and Gastroenterology Nutrition, 20, 333–338. ? ? ? Mart?nez-Villaluenga, C., Fr?as, J., Gomez, R., & Vidal-Valverde, C. (2006). In?uence of addition of ra?nose family oligosaccharides on
F.C. Prado et al. / Food Research International 41 (2008) 111–123 quality vegetable juices enriched with brewer’s yeast autolysate. Food Chemistry, 100, 599–602. Reid, G. (2006). Safe and e?cacious probiotics: What are they? Trends in Microbiology, 14, 348–352. Reid, G., Beuerman, D., Heinemann, C., & Bruce, A. W. (2001b). Probiotic Lactobacillus dose required to restore and maintain a normal vaginal ?ora. FEMS Immunology and Medical Microbiology, 32, 37–41. Reid, G., Bruce, A. W., Fraser, N., Heinemann, C., Owen, J., & Henning, B. (2001a). Oral probiotics can resolve urogenital infections. FEMS Microbiology and Immunology, 30, 49–52. Reid, G., Bruce, A. W., & Taylor, M. (1995). Instillation of Lactobacillus and stimulation of indigenous organisms to prevent recurrence of urinary tract infections. Microecology Therapy, 23, 32–45. Saavedra, J. M., Bauman, N. A., Oung, I., Perman, J. A., & Yolken, R. H. (1994). Feeding of Bi?dobacterium bi?dum and Streptococcus thermophilus to infants in hospital for prevention of diarrhea and shedding of rotavirus. Lancet, 344, 1046–1049. Sanders, M. E. (1998). Overview of functional foods: Emphasis on probiotic bacteria. International Dairy Journal, 8, 341–347. ? Santos, M. C. R. (2001). Desenvolvimento de bebida e farinha lactea ? fermentada de acao probiotica a base de soro de leite e farinha de mandioca ?? por cultura mista de Lactobacillus plantarum A6, Lactobacillus casei Shirota e Lactobacillus acidophilus. MSc thesis. UFPR, 106 pp. Schrezenmeir, J., & De Vrese, M. (2001). Probiotics, prebiotics, and symbiotics – Approaching a de?nition. American Journal of Clinical Nutrition, 73(Suppl.), 361S–364S. Shah, N. P. (2001). Functional foods from probiotics and prebiotics. Food Technology, 55, 46–53. Shanahan, F. (2000). Probiotics and in?ammatory bowel disease: Is there a scienti?c rationale? In?ammatory Bowel Diseases, 6, 107–115. Sheih, Y. H., Chiang, B. L., Wang, L. H., Chuh, L. K., & Gill, H. S. (2001). Systemic immunity enhancing e?ect in healthy subjects following dietary consumption of the lactic acid bacterium Lactobacillus rhamnosus HN001. Journal of the American College of Nutrition, 20, 149–156. Shimakawa, Y., Matsubara, S., Yuki, N., Ikeda, M., & Ishikawa, F. (2003). Evaluation of Bi?dobacterium breve strain Yakult-fermented soymilk as a probiotic food. International Journal of Food Microbiology, 81, 131–136. Shornikova, A. V., Isolauri, E., Burkanova, L., Lukovnikova, S., & Vesikari, T. (1997). A trial in the Karelian Republic of oral rehydration and Lactobacillus GG for treatment of acute diarrhoea. Acta Paediatrica, 86, 460–465. Sieber, R., & Dietz, U. T. (1998). Lactobacillus acidophilus and yogurt in the prevention and therapy of bacterial vaginosis. International Dairy Journal, 8, 599–607. Sodini, I., Lucas, A., Tissier, J. P., & Corrieu, G. (2005). Physical properties and microstructure of yoghurts supplemented with milk protein hydrolysates. International Dairy Journal, 15, 29–35.
Sorenson, D., & Bogue, J. (2005). A conjoint-based approach to concept optimization: Probiotic beverages. British Food Journal, 107, 870–883. Stanton, C., Gardiner, G., Meehan, H., Collins, K., Fitzgerald, G., Lynch, P. B., & Ross, R. P. (2001). Market potential for probiotics. American Journal of Clinical Nutrition, 73(Suppl.), 476S–483S. Stark, A., & Madar, Z. (1994). Dietary ?ber. In I. Goldberg (Ed.). Functional foods: Designer foods, pharmafoods, nutraceuticals (pp. 183–201). New York, EUA: Chapman & Hall. Sultana, K., Godward, G., Reynolds, N., Arumugaswamy, R., Peiris, P., & Kailasapathy, K. (2000). Encapsulation of probiotic bacteria with alginate-starch and the evaluation of survival in simulated gastrointestinal conditions and in yoghurt. International Journal of Food Microbiology, 62, 47–55. Superfoods. (2006). Available at www.livesuperfoods.com (accessed in 11 July 2006). Szajewska, H., Kotowska, M., Mrukowicz, J. Z., Armanska, M., & Mikolajczyk, W. (2001). E?cacy of Lactobacillus GG in prevention of nosocomial diarrhoea in infants. Journal de Pediatria, 138, 361–365. Tuorila, H., & Cardello, A. V. (2002). Consumer responses to an o??avour in juice in the presence of speci?c health claims. Food Quality and Preference, 13, 561–569. Valio. (2006). Evolus? fermented milk drink. Functional products. Available at www.valio.? (accessed in 22 February 2007). Vinderola, C. G., Gueimonde, M., Delgado, T., Reinheimer, J. A., & De ? Los Reyes-Gavilan, C. G. (2000). Characteristics of carbonated fermented milk and survival of probiotic bacteria. International Dairy Journal, 10, 213–220. ? Wacher, C., Canas, A., Barzana, E., Lappe, P., Ulloa, M., & Owens, J. D. ? (2000). Microbiology of Indian and Mestizo pozol fermentation. Food Microbiology, 17, 251–256. Wang, Y.-C., Yu, R.-C., & Chou, C.-C. (2002). Growth and survival of bi?dobacteria and lactic acid bacteria during the fermentation and storage of cultured soymilk drinks. Food Microbiology, 19, 501–508. Wrick, K. L. (1994). The potential role of functional foods in medicine and public health. In I. Goldberg (Ed.). Functional foods: Designer foods, pharmafoods, nutraceuticals (pp. 480–494). New York: Chapman & Hall. Yoon, K. Y., Woodams, E. E., & Hang, Y. D. (2004). Probiotication of tomato juice by lactic acid bacteria. Journal of Microbiology, 42, 315–318. Yoon, K. Y., Woodams, E. E., & Hang, Y. D. (2005). Fermentation of beet juice by bene?cial lactic acid bacteria. Lebensmittel-Wissenschaft und-Technologie, 38, 73–75. Yoon, K. Y., Woodams, E. E., & Hang, Y. D. (2006). Production of probiotic cabbage juice by lactic acid bacteria. Bioresource Technology, 97, 1427–1430. Ziemer, C. J., & Gibson, G. R. (1998). An overview of probiotics, prebiotics and symbiotics in the functional food concept: Perspectives and future strategies. International Dairy Journal, 8, 473–479.