Lactobacillus paracasei-Enriched Vegetables Containing Health Promoting Molecules

Chapter 24

Lactobacillus paracasei-Enriched Vegetables Containing Health Promoting Molecules P. Lavermicocca*, M. Dekker†, F. Russo‡, F. Valerio*, D. Di Venere* and A. Sisto* *Institute of Sciences of Food Production, National Research Council of Italy, Bari, Italy, †Wageningen University, Food Quality and Design Group, Wageningen, The Netherlands, ‡Laboratory of Nutritional Pathophysiology, I.R.C.C.S. “Saverio de Bellis,” National Institute of Digestive Diseases, Bari, Italy

1 INTRODUCTION 1.1  Probiotic Bacteria and Beneficial Effects It is currently recognized that the human microbiota, in particular microorganisms inhabiting the gastrointestinal (GI) tract, has a deep impact on normal physiology and health. Alteration of GI microbiota (dysbiosis) has been associated with the occurrence of a number of diseases, not only of the GI tract itself. Therefore, manipulation of the GI microbiota is considered a potentially suitable approach for maintaining health and preventing and/or treating relevant diseases. In this regard, a marked research and commercial interest has been increasing in the last decades for the development of products containing live probiotic microorganisms which have been defined by FAO and WHO as “live micro-organisms which, when administered in adequate amounts, confer a health benefit on the host” (FAO/WHO, 2001, 2002). Many commercial products, including fermented foods, food complements, and pharmaceutical preparations, contain bacterial strains belonging to Bifidobacterium and Lactobacillus species which are widely used as probiotics. Health benefits which have been claimed for probiotics include, for example, alleviation of intestinal bowel disease symptoms (Orel and Kamhi Trop, 2014), improvement of the normal microflora composition (Cha et al., 2012), pathogen inhibition by producing bactericidal substances and competing with pathogens for adherence to the intestinal epithelium (Liévin-Le Moal and Servin, 2014), reduction of serum cholesterol (Mistry, 2014), prevention of allergies (Enomoto et al., 2014), stabilization and enhancement of the gut mucosal and epithelial barrier function (Rao and Samak, 2013), improvement in the digestion of lactose in intolerant hosts (de Vrese et al., 2001), reduction of constipation symptoms (Yang et al., 2008), and most importantly, modulation of the immune system which can be considered as one of the most important mechanisms responsible for the beneficial effects of probiotic bacteria on human health (Borchers et al., 2009). Finally, an anticarcinogenic activity has also been attributed to probiotics (Kumar et al., 2010); in fact, for example, the regular consumption of probiotics has been associated with the reduction in specific fecal enzymatic activities, including that of β-glucuronidase and β-glucosidase involved in the generation of potentially carcinogenic metabolites in the colon.

1.2  Lactobacillus paracasei as Probiotic L. paracasei, a bacterial species of the L. casei group, has been frequently isolated from foods such as fermented vegetables, milk, dairy products and it is also considered a bacterium that can be found as a common inhabitant of the human intestinal tract. Because of a long history of safe human consumption, this species, together with other lactic acid bacteria, is generally recognized as safe (GRAS status). In fact, although very few and particular strains have been associated with infection cases, the species L. paracasei has been included in the Inventory of Microorganisms with Documented History of Safe Use in Food (Mogensen et al., 2002) as well as in the Qualified Presumption of Safety List by the European Food Safety Authority (EFSA) (Barlow et al., 2007) and a number of selected L. paracasei strains have been used as probiotics. For example, on the basis of DNA analyses, it has been ascertained that L. casei Shirota, a probiotic strain with a very long history of proven health benefits and safe use in the probiotic product Yakult, as well as the L. casei strain contained in

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the well-known yogurt drink, Actimel (Danone) actually belong to the L. paracasei species (Holzapfel et al., 2001). Other L. paracasei strains have been studied as well in relation to their probiotic properties. The patented probiotic strain L. paracasei LMG P-17806 “F19” and the probiotic strain L. paracasei IMPC2.1 (LMG P-22043) (Lavermicocca et al., 2005) were found to be able to induce a low grade pro-inflammatory response, not resulting in an actual inflammatory condition, but enough to induce an enhanced state of host immune system alertness (D’Arienzo et al., 2011). Probiotic activities of L. paracasei IMPC2.1 also include reduction of constipation symptoms (Riezzo et al., 2012), inhibition of the growth of the food-borne human pathogen Yersinia enterocolitica (Lavermicocca et al., 2008), and antiproliferative activity on both gastric and colon cancer cell lines (Orlando et al., 2012). This strain has been recognized by EFSA as sufficiently characterized and identified at the strain level (Sisto et al., 2009). Due to its pro-technological properties, it has been used for the development of innovative and patented probiotic vegetable products whose commercialization has been authorized by the Italian Ministry of Health (Lavermicocca et al., 2003, 2004; Valerio et al., 2013). Interestingly, other L. paracasei strains are currently studied for their remarkable immunomodulatory (anti-inflammatory) activity which is potentially suitable for prevention and/or therapy of diseases in which an inflammatory condition is relevant (D’Arienzo et al., 2011; von Schillde et al., 2012).

1.3 Probiotic L. paracasei and Vegetables Most probiotics available on the market are milk-based foods or dietary supplements, but consumers and enterprises are interested in broadening the range of probiotic food types that can contribute to a regular consumption of probiotics by individuals with lactose intolerance or on cholesterol-lowering diets or for consumers following a diet lacking milk-derived products. To commercialize this market share, producers of probiotic foods are searching to include selected microorganisms with proved probiotic properties in foods and beverages which are part of an everyday diet to provide health benefits while enjoying meals. The intrinsic richness in functional components (such as minerals, vitamins, dietary fibers, antioxidants, and other bioactive molecules) makes fruits and vegetables ideal media for probiotic development (Shahidi, 2009). Particularly, the presence of dietary fibers, whose role in functionality and health is well defined, is responsible for the health perception of plant-based foods by the consumers. However, the survival of strains in non-dairy products is still a challenge that is related to the specific strain endurance and to the intrinsic and extrinsic factors of the food matrix. Recently, Martins et al. (2013) and Rivera-Espinoza and Gallardo-Navarro (2010) reviewed the latest studies on the production of functional foods and particularly probiotic foods obtained by traditional fermentations or by applying innovative technologies (microencapsulation) to several vegetable matrices. Fruits, vegetables, and cereals were processed in various ways to obtain juice, drinks, bars, food mixtures, smoothies, beverages fermented or fortified with probiotic strains (Coda et al., 2012; Nicolesco and Buruleanu, 2010; Rodgers, 2007). Due to the long history of food fermentation, products containing lactic acid bacteria are perceived by the consumers as health-promoting. Probiotic strains used as starter culture may combine the positive image of fermented foods with the functional appeal of the probiotic. Indeed, fortification with a L. paracasei probiotic strain was demonstrated for several vegetable types and extensively verified using table olives, artichokes, and cabbage with the realization of new probiotic vegetable products (Lavermicocca et al., 2005; Sarvan et al., 2013; Valerio et al., 2006). For the development of probiotic table olives, strain L. paracasei IMPC2.1 was selected on the basis of its probiotic properties (see Section 1.2); after that, its suitability as a starter was studied at a laboratory level and then in an industrial plant. Experiments allowed to assess the persistence and the efficacy of strain IMPC2.1 in sustaining the fermentation process, and to ascertain its ability in colonizing the olive surface, persisting in high numbers (more than 7 log10 CFU/g) throughout a shelf-life of more than 3 months (De Bellis et al., 2010). Probiotic olives are considered a functional product because they provide to the consumer more than 9 log10 CFU/serving of the probiotic strain as required by recognized standards for probiotic foods (FAO/WHO, 2001, 2002). Additionally, with regard to the organoleptic profile, the process allows obtaining a final product submerged in 4% NaCl brine (a NaCl concentration lower than the usual 8–9%) with improved flesh texture and flavors. From a nutritional point of view, probiotic olives (cv. Bella di Cerignola) are low caloric (103 kcal/100 g), and contain 2.9% of fibers, 0.2% of carbohydrates, and 11% of fats (P. Lavermicocca, Unpublished results). Even though information on fat composition in table olives is scarce, an interesting investigation by Lopez et al. (2006) revealed that green table olives contain monounsaturated fatty acids as the main components of total fat, while saturated fatty acids were, in general, fairly low. Furthermore, table olives can be considered “functional” for the presence of significant amount of selenium, an essential element for the human organism. In fact, among vegetables and fruits, olives are one of the main sources of selenium (on average 22 ng/g) (Diaz-Alarcón et al., 1994).

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Table olives were validated as probiotic food for transporting bacterial cells into the human GI tract. Actually, L. paracasei IMPC2.1 was recovered from fecal samples of volunteers fed 10–15 olives per day carrying about 9 log10 viable cells for 10 days (Lavermicocca et al., 2005). Besides table olives, artichokes preserved in brine in the presence of the strain sustained viable probiotic bacterial populations in the product during storage. Artichokes were subjected to a mild cooking step to limit enzymatic activities and then soaked in a brine solution containing L. paracasei IMPC2.1 in order to obtain a final concentration of about 8 log10 CFU of live probiotic cells per gram (Valerio et al., 2006, 2013). Artichokes were then lightly seasoned with olive oil, packed in modified atmosphere and stored under refrigeration. The probiotic strain was able to survive for at least 3 months indicating that artichokes are a suitable food supporting the bacterial survival during storage and therefore can deliver about 9–10 log10 live cells per portion (consisting of about 100 g). As observed for table olives, also in the case of artichokes, the culture of L. paracasei IMPC2.1 may act in a double role of probiotic and as a bioprotective culture (Valerio et al., 2013). Biopreservation is an approach that has gained attention in recent years and can be defined as the extension of shelf life and food safety by the use of natural or controlled microbiota and/or their antimicrobial compounds (Stiles, 1996). Recent investigations have recognized fruits and vegetables as vehicles for the transmission of human pathogens; actually an increase of the incidence of foodborne outbreaks caused by contaminated fresh fruits and vegetables has been recently reported (Berger et al., 2010). For acidified ready-to-eat vegetable- or fruit-based products (foods to which acids have been added), even if the risk of pathogens’ growth is considered not significant due to the low pH value (below 4.6), pathogenic microorganisms can adapt to acidic conditions and survive during shelf life (Gandhi and Chikindas, 2007; Uyttendaele et al., 2009). Valerio et al. (2013) observed that the probiotic L. paracasei IMPC2.1 ensured the safety of artichokes during 2 months (product shelf life) under refrigerated storage because of its antagonistic activity against the pathogens Listeria monocytogenes, Salmonella enterica subsp. enterica, and Escherichia coli avoiding a pasteurization step. Therefore, the probiotic strain, representing always more than 93% of lactic acid bacteria (about 7 log10 CFU/g) during the entire shelf life is also active as a protective culture. The strain can combine the controlling of the development of pathogens during storage with the probiotic benefits and is suitable for industrial processing of vegetables. Moreover, the probiotic L. paracasei IMPC2.1 was further successfully applied to cabbage fermentation leading to a functional food characterized by a high content of bioactive molecules and live probiotic bacterial cells (Sarvan et al., 2013) (see Section 2). In conclusion, the use of probiotics allows a mild preservation technique satisfying consumer demand for more natural and fresh-like foods and meets the latest trends toward traditional foods with additional functional benefits.

2  L. PARACASEI-ENRICHED CABBAGE AS SOURCE OF HEALTH-PROMOTING PHYTOCHEMICALS AND CARRIER OF PROBIOTIC CELLS Cabbage (Brassica oleracea var. capitata) is a cruciferous vegetable that is globally widely consumed in the human diet in many cultures. The Brassicaceae family is rich in phytochemicals such as polyphenols, carotenoids, and glucosinolates (GSs). For GSs they are virtually the only source within the human diet. A diet rich in vegetables of the family Brassicaceae has been shown in epidemiological studies to reduce the risk of several types of cancers (Kim and Park, 2009; Steinbrecher et al., 2009). GSs are reported to be involved in the protection of the human body against several stages in the development of cancers (Verkerk et al., 2009). The basic chemical structure of GSs is a β-thioglucosideN-hydroxysulfate with a sulfur linked β-d-glucopyranose moiety and a side chain, which is either aromatic, indolic, or aliphatic. The GS content and profile are different in different brassica varieties, but depend also on agronomic practices and conditions (Verkerk et al., 2009). The most common GSs found in white cabbage are glucoiberin, progoitrin, sinigrin, glucobrassicin, and 4-methoxyglucobrassicin (Castro et al., 2004; Ciska et al., 2000). If GSs are broken down enzymatically, different products such as isothiocyanates, nitriles, thiocyanates, and epithionitriles can be released depending on the environmental conditions of the reaction (Verkerk et al., 2009). Isothiocyanates are assumed to be bioactive and responsible for the health benefits of Brassicaceae (Traka and Mithen, 2009; Verkerk et al., 2009). Researches to establish the mechanisms leading to these protective effects have been carried out (Mithen et al., 2000; Verkerk et al., 2009), and for the most prominent isothiocyanate, i.e. sulforaphane, the following mechanisms are reported: induction of detoxification enzymes to convert and excrete carcinogens, induction of cell cycle arrest and apoptosis, inhibition of angiogenesis and metastasis, changes in histone acetylation status, and antioxidant, antiinflammatory, and immunomodulatory activities (Dinkova-Kostova and Kostov, 2012).

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Enzymatic breakdown of GSs can take place during domestic preparation, industrial processing, or chewing, when the intact cell tissue is destroyed. Damaged plant tissues allow the endogenous enzyme myrosinase (β-thioglycosidase, E.C. 3.2.1.147), which is stored separately from the GSs in intact plant tissue (Kissen et al., 2009), to hydrolyse GSs. If GS containing food is ingested without active myrosinase being present (e.g., due to cooking), up to 40% of ingested GSs can be hydrolyzed by the human gut flora (Fahey et al., 2012) and the bioactive breakdown products are absorbed by the colon. Besides by enzymatic hydrolysis, GS levels can also be affected by industrial or domestic processing involving heat treatments of Brassica vegetables due to thermal degradation (Oerlemans et al., 2006) and leaching into the processing solutions. A mathematical model to predict the retention of phytochemicals during processing of vegetables has been developed and applied to estimate the content of GSs in Brassica after thermal treatment. This model can help optimizing the health benefits of vegetable products for consumers (Sarvan et al., 2012). For the traditional fermentation of cabbage to sauerkraut, the raw vegetable is sliced into thin strips and placed in a brine solution. From the damaged vegetable cells, GSs as well as myrosinase, can diffuse and the enzyme can get into contact with GSs and will hydrolyse them (Verkerk et al., 2009). This has been shown to be detrimental for the amounts of GSs in the final fermented products (Martinez-Villaluenga et al., 2009; Tolonen et al., 2002). In order to develop a fermented cabbage still containing GSs, it seems necessary to inactivate the myrosinase activity prior to the fermentation process. A thermal treatment like a mild blanching step can be used for this. Due to such a heat treatment the naturally present microbial flora will also be disrupted. Therefore, a starter culture should be added to initiate the fermentation process after blanching. Actually, the probiotic L. paracasei IMPC2.1 has been used for this having the advantage that not only the GS content will be retained, but also the probiotic can add health benefits to the final fermented food (Sarvan et al., 2013). It was shown that blanched white cabbage supports the growth of the strain which performs an appropriate acidification, persisted during refrigerated vacuum-packed storage, and prevented vegetable deterioration, thereby being suitable for industrial processing. The probiotic strain was able to grow in the blanched cabbage without nutrient supplementation reaching counts of about 8 log10 CFU/g in brine (4% NaCl) replacing the background flora. An efficient acidification occurred in probiotic cabbage even though a low inoculum load (about 4 log10 CFU/g), 1000 times lower than that usually used in industrial practices for vegetable fermentation, was applied. The probiotic population remained steady (about 8 log10 CFU/g) during the 30-day vacuum package storage period at 4 °C determining a desirable sensory quality of the product. Figure 24.1 shows the total GSs content after the different processing steps: raw, blanched, and fermented by the probiotic starter, with respect to the content in traditional sauerkraut (Sarvan et al., 2013). The content of GSs was reduced by about 50% due to the blanching step (100 °C, 5 min): the GS loss during blanching will be mainly due to leaching into the blanching water. These losses can be reduced by a shorter blanching time and use of steaming instead of boiling water for the blanching. The probiotic fermentation reduced the GSs content by 35% compared to the blanched cabbage while in traditional sauerkraut no GSs are detected after the fermentation process. So the pre-treatment of the cabbage prior to the fermentation resulted in a drastic improvement of the GSs content of the final product. Vacuum packaging and storage of the obtained fermented product for 30 days resulted in very minor (13%) reduction of the GSs content indicating that the compound is stable during the product shelf life (Sarvan et al., 2013). It can be concluded that the probiotic cabbage is far superior above traditional sauerkraut, even though further improvement in the content can still be obtained by optimizing the blanching and fermentation step.

FIGURE 24.1  GSs content during the processing and storage and compared with the traditional product.

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3  L. PARACASEI-ENRICHED ARTICHOKES AS A SYMBIOTIC 3.1  The Artichoke as a Source of Bioactive Compounds The growing interest for foods rich in “nutraceuticals” has directed the research attention to the artichoke, due to its high content in inulin and polyphenols, two classes of compounds with prebiotic and antioxidant properties, respectively. Prebiotic molecules selectively stimulate the growth of probiotic bacteria in the colon. The addition of prebiotic molecules is a widespread strategy to improve the functional quality of probiotic foods. Frequently, dairy products, especially yogurt, are enriched with dietary fibers, for example inulin. Inulin and other nondigestible carbohydrates (oligo- and polysaccharides), because of their chemical structures, are not hydrolyzed by human digestive enzymes and not absorbed in the upper part of the GI tract. Such molecules could be called “colonic foods,” i.e. foods entering the colon and serving as substrates for the endogenous colonic bacteria, thus indirectly providing the host with energy, metabolic substrates, and essential micronutrients (Gibson and Roberfroid, 1995). Inulin, a polymer of fructose also present in other vegetable species (garlic, onion, Jerusalem artichoke, chicory), is classified as soluble fiber and represents the storage polysaccharide of the plant. Its polymerization degree principally depends on genetic factors and can be influenced by agronomic and physiological factors. The average polymerization degree of artichoke inulin is about 30 and 20 in roots and heads, respectively (D. Di Venere, 2014, unpublished data). The inulin content of an artichoke head depends on many factors, i.e. genotype, environment, physiological stage, and time of harvesting (Di Venere et al., 2005b). During the artichoke head growth, inulin content significantly increases, ranging from about 8% f.w. (43% d.w.) in wintertime to 5.5% f.w. (35% d.w.) in springtime (Di Venere et al., 2005a). The interest in potential medical uses of inulin and inulin derivatives is growing, because it was demonstrated that a fructan-rich diet may have health-promoting effects (Causey et al., 2000). Actually, dietary fructans are fermented in the large intestine resulting in an increase in microbial mass and production of short-chain fatty acids (SCFAs) (Wang and Gibson, 1993). Inulin has been shown to favor the growth of indigenous lactobacilli and/or bifidobacteria and to improve the functionality of the GI system (Roberfroid, 2002; Gibson and Roberfroid, 1995). For this reason, artichokes were chosen as suitable vegetable matrix for the development of a probiotic product; in fact, inulin contributed to an efficient implantation of the probiotic L. paracasei IMPC2.1 (Valerio et al., 2006) (see Section 3.2). As a beneficial effect of inulin and oligofructose on lipid metabolism, some studies have shown that a diet supplemented with 10 g/day inulin for 8 weeks can significantly decrease blood triglyceride levels (Izzo and Franck, 1998). Furthermore, a positive action of dietary inulin and oligofructose on the modification of cancer risk has been shown (Pool-Zobel, 2005; Taper and Roberfroid, 2002). Besides inulin, the “nutraceutical” properties of artichokes are attributed to the presence of polyphenols, in particular hydroxycinnamic acid derivatives and flavonoids (apigenin and luteolin glycosides). Caffeoylquinic acids are the most predominant artichoke head phenolic compounds, the most abundant being chlorogenic acid (5-O-caffeoylquinic acid) and two dicaffeoylquinic acids (1,5-O- and 3,5-O-dicaffeoylquinic acid). Phenolic content is influenced by genetic, physiological, and environmental factors and significantly changes with genotype, physiological stage, and age of artichoke field. Polyphenol content in the edible part of the head ranges between 0.5% and 2% f.w. (about 3–12% d.w.) depending on cultivars (Di Venere et al., 2005a,b). Azzini et al. (2007) studied the absorption and metabolism of phenolic derivatives in human plasma after ingestion of cooked artichokes. Moreover, recent studies have shown the antioxidant and apoptotic activity of phenolic compounds extracted from the edible part of the artichoke head in rat hepatocytes in primary cell culture and human hepatoma cell lines (Miccadei et al., 2008). Also, the apoptotic activity of such polyphenolic extracts on a human breast cancer cell line was demonstrated (Mileo et al., 2012).

3.2  Probiotic Artichokes and GI Function Artichokes fortified with the probiotic L. paracasei IMPC2.1 have been tested in constipated and healthy human subjects to assess the efficacy of this symbiotic food in modulating the gut flora and in alleviating symptoms of functional constipation.

3.2.1  L. paracasei-Enriched Artichokes Modulates Gut Microbial Parameters in Humans An important factor influencing probiotic efficacy is the ability of the strain to survive food processing and storage and, after human consumption, the GI tract. The food carrier should contain functional ingredients that positively interact with the probiotic, sustaining its survival and growth in the food product, buffering bacterial cells through the GI tract and regulating their colonization (Ranadheera et al., 2010). Experiments carried out to demonstrate the protective action of artichokes and olives showed that a high percentage of the L. paracasei IMPC2.1 population anchored to vegetables survived during

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simulated gastric and intestinal digestion (Valerio et al., 2006). The high recovery rate of the total bacterial population anchored to artichokes or olives or suspended in skim milk could be explained by the protective action of the fat content for olives and milk or to the presence of prebiotic carbohydrates for artichokes. Actually, the presence of prebiotic substances in the food matrix, such as inulin in artichokes, can further strengthen the probiotic efficacy into the human gut. Dietary components, in particular carbohydrates not digested in the upper gut (see Section 3.1), are metabolized by “beneficial bacteria” in the large intestine where they are transformed to lactic acid, hydrogen, carbon dioxide, and SCFAs and then rapidly adsorbed (Bekkali et al., 2007; Brouns et al., 2002; Cummings et al., 1987). The three major SCFAs are acetate, propionate, and butyrate, and the latter is supposed to maintain gut integrity and to prevent the bacterial translocation occurring in several GI diseases (Perez Chaia and Oliver, 2003). A preliminary in vivo post-test trial on eight human subjects suffering from constipation suggested the efficacy of L. paracasei IMPC2.1 carried by ready-to-eat artichokes (Figure 24.2) to transiently colonize the human gut, thus modulating at the individual level potentially harmful bacteria, fecal enzyme activity, and SCFA production as well as symptom profile (Valerio et al., 2010). This study has been followed by a human trial on 20 healthy subjects consuming L. paracaseienriched ready-to-eat artichokes in comparison to control ready-to-eat artichokes (Valerio et al., 2011). The study demonstrated that the probiotic strain transiently colonized the gut of 85% of subjects, caused the inhibition of potentially harmful bacteria, and stimulated the growth of LAB and of presumptive lactobacilli and bifidobacteria. Whereas for subjects enrolled in control group, a different microbial behavior was observed with a trend to the increase of potentially harmful bacteria and in particular of E. coli counts. Interestingly, the Shannon diversity index confirmed a higher genetic diversity of LAB only in the probiotic group with respect to the start of the study while this index remained almost unvaried in the control group. A slight increase of total SCFA concentrations was observed in the probiotic group in comparison to the start of the study. Generally, only the consumption of probiotic artichokes during the first 15-day period caused a shift of microbial counts toward lower values of potentially harmful bacteria (Enterobacteriaceae, E. coli, total Clostridium spp.) and higher values of LAB and of presumptive lactobacilli and bifidobacteria. Most recently the efficacy of a L. paracasei strain DG in modulating the intestinal microbial ecology has been evaluated in a randomized double-blind, placebo-controlled crossover trial on 34 healthy human volunteers (Ferrario et al., 2014). The study demonstrated that despite the high inter-individual variability in microbiota composition, the 4-week consumption of the probiotic strain modified the local microbial ecology with particular reference to Clostridiales populations, and caused a rebalancing effect on SCFA concentrations strictly dependent on the initial microbial ecosystem. Additionally, Zhang et al. (2013) assessed the probiotic efficacy of the L. paracasei subsp. paracasei LC01 in a randomized, double-blind, placebo-controlled human trial and observed a significant inhibition in fecal E. coli, increase in Lactobacillus, Bifidobacterium, and Roseburia intestinalis and in acetic and butyric acids.

3.2.2  Possible Clinical Applications Current trends for probiotics in GI diseases include the possible management of some disturbances such as gastric and colonic neoplasms, intestinal inflammatory diseases, postoperative complications, acute GI infection, diarrhea, and lactose

FIGURE 24.2  Ordinary and probiotic artichokes packed in identical trays to obtain ready-to-eat artichoke products (about 180 g).

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intolerance. Interestingly, disturbances in the gut microbiota may contribute to symptomatology and even, the etiology of functional diseases such as functional constipation (Quigley, 2008). Among GI nonorganic diseases, functional constipation is caused by nonorganic or drug factors. It is a common problem in westernized societies and is characterized by abdominal discomfort or pain, abdominal distension, headache, dizziness, and poor appetite, all symptoms that can interfere heavily with the quality of life of patients (Higgins and Johanson, 2004). Different therapeutic strategies have been hypothesized for fighting functional constipation. A new therapeutic approach for constipation could be based on modulating intestinal microflora which can influence peristalsis of the colon. Currently, probiotics are increasingly being used in the management of this condition. There are several reasons why probiotics might have therapeutic potential for the treatment of constipation (Guarner and Malagelada, 2003). Firstly, there are data demonstrating differences in the intestinal microbiota between healthy individuals and patients with chronic constipation. Secondly, studies involving the administration of Bifidobacterium lactis DN-173 010 have shown improved colonic transit times, both in a healthy population and in constipated patients (Picard et al., 2005). Finally, probiotics lower the pH in the colon. This reduction in pH is due to the bacterial production of SCFAs (butyric acid, propionic acid, and lactic acid). A lower pH enhances peristalsis in the colon and, subsequently, might decrease the colonic transit time (Chmielewska and Szajewska, 2010). Probiotic-enriched ready-to-eat artichokes represent a gastronomic preparation that ensure the long lasting surviving of probiotics. In this framework, Riezzo et al. (2012) evaluated the effects of probiotic-enriched artichokes on treatment preference, symptom profile, and SCFA production in constipated subjects when compared with control ready-to eat artichokes. The study was performed on constipated patients as a double-blind, placebo-controlled, crossover randomized clinical trial in Castellana G., Italy. A group of patients (n 20) suffering from functional constipation was studied using a double-blind method and a computer-generated randomization list. Each patient consumed 180 g per day of ordinary artichokes or artichokes enriched with L. paracasei IMPC2.1 for 15 days (daily dose of 20 log10 CFU). Ordinary and probiotic artichokes were lightly seasoned with olive oil and packed in identical trays with modified atmosphere to obtain ready-to-eat artichoke products. Final products had identical shape, texture, and appearance and there was no way to distinguish between the two products (Figure 24.2). Relief of symptoms was evaluated by a visual analogue scale. The stool consistency and symptom profile of subjects were investigated by Bristol stool form chart and the Gastrointestinal Symptom Scoring Rate questionnaire (GSRS). At the end of the study, 80% of patients preferred probiotic-enriched artichokes to ordinary ones. As concern symptom profile, satisfactory relief of symptoms was significantly higher during the probiotic-enriched artichoke period. Additionally, significant increase in Bristol score, along with significant reduction in GSRS constipation cluster score and in its single items (frequency of evacuation, hard stools, and feeling of incomplete evacuation), were observed. A variety of factors including probiotic strain, dose, and treatment duration, and adequate vehicle may affect therapeutic outcome in the management of constipation by administering probiotics. In this study (Riezzo et al., 2012), administration of enriched artichokes lasted 15 days and L. paracasei IMPC2.1 was found in the feces of 17 out of 20 patients, thus indicating that duration of probiotic administration was sufficient to permit colonization, without significantly affecting the compliance. Vegetables such as artichokes may act as “active” vehicles for the transport and release of probiotic bacterial populations (Bundy et al., 2004; Riezzo et al., 2012). Besides, the released amounts of probiotic bacteria are comparable or even higher than those of milk-based probiotic products available on the market. In conclusion, the obtained results allow us to suppose that the introduction of some vegetables enriched with probiotics such as artichokes, but also salads and olives, could increase the choices for patients/consumers on a modified dietary regimen by contemporarily amplifying therapeutic effects for GI diseases in a synergistic manner.

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