Safety and functional aspects of pre-selected lactobacilli for probiotic use in Iberian dry-fermented sausages

The purpose of this study was to investigate lactobacilli for potential probiotic use in Iberian dry-fermented sausages. A total of 18 strains, previously selected, isolated from different sources were evaluated according to safety and functional characteristics including biogenic amines and d-lactic acid production, antibiotic susceptibility, haemolysis, cell adhesion, and antimicrobial activity against food-borne pathogens. The strains Lactobacillus reuteri PL519, L. reuteri PL542, and Lactobacillus fermentum HL57 were able to establish their self on the intestinal epithelium, and an inhibition of such pathogenic bacteria as Listeria monocytogenesin vitro. These strains were also considered safe to be used with regard to their low aminogenic potential and d-lactic acid production, antibiotic resistance pattern and haemolytic activity. Thus, they were selected as a potential probiotic meat starter culture suitable for manufacture of dry-fermented Iberian sausages.     

Lactic acid starter and probiotic bacteria: a comparative “in vitro” study of probiotic characteristics and biological barrier resistance

Probiotic characteristics (deconjugation of bile salts, hydrophobicity and β-galactosidase activity) and the resistance to biological barriers (gastric juice and bile salts) of 24 strains of lactic acid starter bacteria (Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus and Lactococcus lactis) and 24 strains of probiotic bacteria (Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus rhamnosus and bifidobacteria) were compared. Among the probiotic bacteria tested, Lactobacillus acidophilus was the most interesting species since it showed high values of resistance to gastric juice and bile, hydrophobicity and β-galactosidase and bile salts deconjugation activities. Bifidobacterium bifidum strains showed the same behavior, although the values of the parameters investigated were slightly lower than those obtained for Lactobacillus acidophilus. On the other hand, it was demonstrated that Lactobacillus delbrueckii subsp. bulgaricus was the lactic acid starter species with the best probiotic characteristics among the starter species assessed. It was resistant to gastric juice and bile, and showed high values for β-galactosidase activity. On the other hand, lactic acid starter bacteria showed hydrophobicity values similar to or higher than those obtained for the strains of the Lactobacillus casei . According to the results found, the total probiotic value of a fermented dairy product should take into account not only the intestinal probiotic cultures used in the formulation but also the probiotic contribution of the lactic acid starter microflora.     

In vitro fermentation of cereal dietary fibre carbohydrates by probiotic and intestinal bacteria

A range of probiotic and other intestinal bacteria were examined for their ability to ferment the dietary fibre carbohydrates β‐glucan, xylan, xylo‐oligosaccharides (XOS) and arabinoxylan. β‐Glucan was fermented by Bacteroides spp and Clostridium beijerinckii but was not fermented by lactobacilli, bifidobacteria, enterococci or Escherichia coli. Unsubstituted xylan was not fermented by any of the probiotic bacteria examined. However, many Bifidobacterium species and Lactobacillus brevis were able to grow to high yields using XOS. XOS were also efficiently fermented by some Bacteroides isolates but not by E coli, enterococci, Clostridium difficile, Clostridium perfringens or by the majority of intestinal Lactobacillus species examined. Bifidobacterium longum strains were able to grow well using arabinoxylan as the sole carbon source. These organisms hydrolysed and fermented the arabinosyl residues from arabinoxylan but did not substantially utilise the xylan backbone of the polysaccharide. Arabinoxylan was not fermented by lactobacilli, enterococci, E coli, C perfringens or C difficile and has potential to be an applicable carbohydrate to complement probiotic Bif longum strains in synbiotic combinations. © 2002 Society of Chemical Industry     

Probiotic potential of Lactobacillus delbrueckii strains and their phage resistant mutants

Three commercial phage sensitive strains of Lactobacillus delbrueckii and spontaneous phage resistant mutants isolated from them were studied by focusing on their biological and probiotic features. After incubation in a simulated gastric solution (pH 2.0), viable cell counts decreased moderately, but a protective effect on strains was observed when milk was added. A limited resistance to bile was observed, while most of the strains tolerated lysozyme, and grew in the presence of bile salts and fermented prebiotics. Some strains showed high hydrophobicity values and β-galactosidase activity. The strong antibacterial activity displayed toward pathogens was due to the production of lactic acid. Sensitive strains and their phage resistant variants were able to adhere to Caco-2/TC-7 monolayers and significantly inhibited the invasion of Salmonella enterica serovar Enteritidis into Caco-2/TC-7 cells. All strains tested showed potential probiotic features, suggesting a potential probiotic role of L. delbrueckii.     

Short communication: Development of a direct in vivo screening model to identify potential probiotic bacteria using Caenorhabditis elegans

Caenorhabditis elegans is an accepted model host to study host-bacteria interactions in the gut, in addition to being a simple model with which to study conserved aspects of biological signaling pathways in intestinal environments, because these nematode worms have similar intestinal cells to those of humans. Here, we used C. elegans to develop a new in vivo screening system for potential probiotic lactic acid bacteria (LAB). Initially, critical colonization ability of LAB strains isolated from Korean infant feces was screened in the worm intestinal tract over a period of 5 d. Furthermore, we investigated host health-promoting activities, including longevity-extending effects and immune-enhancing activities against foodborne pathogen infection. We identified 4 LAB strains that were highly persistent in the nematode gut and that significantly prolonged the longevity of C. elegans and improved the survival of C. elegans in response to infection by Staphylococcus aureus. The 4 LAB strains we identified showed resistance to acid and bile conditions, assimilated cholesterol, and were able to attach to a mucus layer. The 4 LAB isolates were identified as Lactobacillus plantarum using 16S rRNA sequencing analysis. Taken together, we developed a direct in vivo screening system using C. elegans to study host health-promoting LAB. Our system is simple, rapid, cost-effective, and reliable, and we anticipate that this system will result in the discovery of many more potential probiotic bacteria for dairy foods.     

Novel probiotic candidates for humans isolated from raw fruits and vegetables

This study was aimed at determining the probiotic potential of a large number of autochthonous lactic acid bacteria isolated from fruit and vegetables. Survival under simulated gastric and intestinal conditions showed that 35% of the strains, mainly belonging to the species Lactobacillus plantarum maintained high cell densities. Selected strains did not affect the immune-mediation by Caco-2 cells. All strains stimulated all 27 immune-mediators by peripheral blood mononuclear cells (PBMC). A significant (P < 0.05; P < 0.01) increase of the major part of cytokines and growth factors was found. A few chemokines were stimulated. Immune-mediators with pro-inflammatory activity (IL-17, EOTAXIN and IFNγ) were significantly (P < 0.01) stimulated by all strains, followed by IL-1b > IP-10 > IL-6 > MIP1α. Stimulation of IL-12, IL-2 and IL-7 was strain dependent. Only a few strains increased the synthesis of cytokines with anti-inflammatory activity. Six L. plantarum strains were further selected. Four were defined as the strongly adhesive strains (more than 40 bacteria adhering to one Caco-2 cell), and 2 as the adhesive strains (5–40 bacteria adhering to one Caco-2 cell). Five strains grew and acidified chemically defined medium with fructo-oligosaccharides (FOS) as the only carbon source. End-products of FOS fermentation were found. All strains inhibited enterohemorragic Escherichia coli K12 and Bacillus megaterium F6 isolated from human sources. The results of this study showed that some autochthonous lactic acid bacteria from raw fruit and vegetables have functional features to be considered as novel probiotic candidates.     

Diversity and probiotic potentials of lactic acid bacteria isolated from gilaburu,a traditional Turkish fermented European cranberrybush (Viburnum opulus L.) fruit drink

The aim of the present study was to characterize lactic acid bacteria (LAB) strains isolated from traditional fermented gilaburu fruit juice and their probiotic potential. The LAB counts of the fermented gilaburu fruit juice were in the range of 3.92–8.30 log cfu/g. Total of 332 isolates belonging to Lactobacillus and Leuconostoc species were characterized from traditional fermented gilaburu juice by genotypic methods. It was also determined that the major LAB strains belong to Lactobacillus plantarum (173 isolates), Lactobacillus casei (52 isolates) and Lactobacillus brevis (24 isolates), while Lactobacillus buchneri, Lactobacillus parabuchneri, Lactobacillus pantheris, Leuconostoc pseudomesenteroides and Lactobacillus harbinensis were the least in isolated LAB strains. In terms of the probiotic potentials, Lb. plantarum strains were able to grow at pH 2.5, but 3 of Lb. casei strains, one of each Lb. brevis and Lb. buchneri strains could not grow at the same pH. All selected LAB stains were resistant to bile salt at ≤ 0.3% concentration. While all the LAB species grew at 15 °C, two Lactobacillus hordei strains could also grow at 45 °C. The highest cell hydrophobicity degrees were for Lb. casei (G20a) and Lb. plantarum (G19e) as 87.5 and 86.0%, respectively. Listeria monocytogenes and Bacillus cereus were the most sensitive bacteria against the selected LAB strains, while Escherichia coli and Staphylococcus aureus were the most resistant. Again all the isolated LAB species were resistant to three antibiotics; kanamycin, streptomycin and vancomycin. Characterization and probiotic potentials of the LAB isolated from fermented gilaburu (Viburnum opulus) juice were studied first time, and further research needs to be done on their behaviors in similar food formulations as a probiotic.     

酵素源乳酸菌的益生功能及抑菌特性评价

为筛选到发酵性能稳定且高效的功能性菌株,以韶关紫背桃源生产的酵素中分离出的12株乳酸菌为试验菌株,测定其益生特性。分别对12株乳酸菌进行了自聚集性、耐酸性,以及对胆盐、胃蛋白酶、胰蛋白酶与抗生素的耐受性和对肠道致病菌的拮抗作用实验,结果表明：12株受试乳酸菌均表现出良好的自聚集性;除XZJ07外,其他11株菌株在pH值为2.5的模拟胃液中培养3 h存活率仍达75%以上;在人工模拟胃液以及肠液中对胃蛋白酶和胰蛋白酶孵育2、3 h存活率仍超过75%;在含有胆盐浓度为3 g/L MRS培养基中培养3 h后,活菌数仍达106 CFU/mL,另外,受试菌株对金黄色葡萄球菌、大肠杆菌均有一定的抑制作用,对大肠杆菌抑菌圈直径均大于15 mm,对金黄色葡萄球菌抑菌圈直径均大于17 mm;受试菌株对氨苄青霉素敏感,对四环素、红霉素、土霉素较为耐受。研究结果预示着受试菌株具有作为优质益生性发酵剂的潜在能力。     

Evaluation of probiotic characteristics of newly isolated Lactobacillus spp.: immune modulation and longevity

In the current study, the probiotic potential of approximately 350 strains of lactic acid bacteria isolated from Korean infant feces and Kimchi was investigated. Common probiotic properties of the bacterial strains, such as acid tolerance, bile tolerance and adhesion to human intestinal epithelial cells (HT-29 cells), were examined. Some strains were found to have immune modulatory and antimicrobial properties. Antagonistic activity against a panel of pathogenic bacteria was found to be strain dependent. To evaluate the immune modulatory activity of the strains, lymphocyte interferon (IFN)-γ secretion was determined in conjunction with cell proliferation. Some strains of Lactobacillus gasseri, L. fermentum and L. plantarum exhibited increased IFN-γ levels and lymphocyte proliferation. To evaluate the effects of these immune modulating lactobacilli on host life span, Caenorhabditis elegans was used as an in vivo model. Nematodes that were supplied heat-killed lactobacilli as a food source exhibited obvious differences in life span compared with those fed Escherichia coli OP50. The mean life span (determined as mean percent survival) of worms fed L. plantarum CJLP133 and L. fermentum LA12 was 13.89% and 13.69% greater, respectively, than that of control nematodes after 21 days (P = 0.036 and 0.043, respectively). In addition, some of safety profiles, including hemolytic type, gelatin hydration and degradation of urea, were found to be positive. These newly identified lactobacilli hold promise for use as probiotic agents, feed additives and/or in food applications.     

Production of exopolysaccharides by Lactobacillus and Bifidobacterium strains of human origin, and metabolic activity of the producing bacteria in milk

This work reports on the physicochemical characterization of 21 exopolysaccharides (EPS) produced by Lactobacillus and Bifidobacterium strains isolated from human intestinal microbiota, as well as the growth and metabolic activity of the EPS-producing strains in milk. The strains belong to the species Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus vaginalis, Bifidobacterium animalis, Bifidobacterium longum, and Bifidobacterium pseudocatenulatum. The molar mass distribution of EPS fractions showed 2 peaks of different sizes, which is a feature shared with some EPS from bacteria of food origin. In general, we detected an association between the EPS size distribution and the EPS-producing species, although because of the low numbers of human bacterial EPS tested, we could not conclusively establish a correlation. The main monosaccharide components of the EPS under study were glucose, galactose, and rhamnose, which are the same as those found in food polymers; however, the rhamnose and glucose ratios was generally higher than the galactose ratio in our human bacterial EPS. All EPS-producing strains were able to grow and acidify milk; most lactobacilli produced lactic acid as the main metabolite. The lactic acid-to-acetic acid ratio in bifidobacteria was 0.7, close to the theoretical ratio, indicating that the EPS-producing strains did not produce an excessive amount of acetic acid, which could adversely affect the sensory properties of fermented milks. With respect to their viscosity-intensifying ability, L. plantarum H2 and L. rhamnosus E41 and E43R were able to increase the viscosity of stirred, fermented milks to a similar extent as the EPS-producing Streptococcus thermophilus strain used as a positive control. Therefore, these human EPS-producing bacteria could be used as adjuncts in mixed cultures for the formulation of functional foods if probiotic characteristics could be demonstrated. This is the first article reporting the physicochemical characteristics of EPS isolated from human intestinal microbiota.     

Functional properties of Lactobacillus strains isolated from kimchi

The objective of this study was to evaluate the functional properties of lactic acid bacteria (LAB) from kimchi, a traditional Korean fermented vegetable product generally consumed raw as a side-dish with practically every meal.Twelve mild acid producing facultatively heterofermentative Lactobacillus strains were selected for their potential as starter cultures for fermentation of kimchi, and evaluated for their functional properties. Eleven strains were identified as Lactobacillus sakei and one as Lactobacillus plantarum. The strains identified as L. sakei differed in some physiological features; of particular interest was the fact that 9 of these strains produced L(+) lactic acid from glucose in presence of acetate.All strains were able to survive gastrointestinal conditions simulating stomach and duodenum passage. In addition, they showed higher adherence to HT-29 cells than Lactobacillus rhamnosus GG, a commercial probiotic strain used worldwide. These strains also showed antimicrobial activity against a number of food-borne pathogens. Their ability to lower cholesterol was demonstrated by BSH (bile salt hydrolytic) activity, and cholesterol assimilation tests in vitro. The results suggest the probiotic potential of these strains for use in kimchi fermentation.     

Influence of cofermentation by amylolytic Lactobacillus strains and probiotic bacteria on the fermentation process,viscosity and microstructure of gruels made of rice,soy milk and passion fruit fiber

Gruels tailored to school-age children and made of soy milk and rice flour with or without total dietary fiber from passion fruit by-product were fermented by amylolytic lactic acid bacteria strains (Lactobacillus fermentum Ogi E1 and Lactobacillus plantarum A6), by commercial probiotic bacteria strains (Lactobacillus acidophilus L10, Lactobacillus casei L26 and Bifidobacterium animalis subsp. lactis B94) and by co-cultures made of one amylolytic and one probiotic strain. The influence of ingredient composition and bacterial cultures on kinetics of acidification, α-amylase activity of the bacteria, apparent viscosity and microstructure of the fermented products was investigated. During fermentation of the gruels, α-amylase activity was determined through the Ceralpha method and apparent viscosity, flux behavior and thixotropy were determined in a rotational viscometer. Rheological data were fitted to Power Law model. The combination of amylolytic and probiotic bacteria strains reduced the fermentation time of the gruels as well as increased the α-amylase activity. The addition of passion fruit fiber exerted less influence on the apparent viscosity of the fermented products than the composition of the bacterial cultures. Scanning electron microscopy provided evidence of exopolysaccharide production by amylolytic bacteria strains in the food matrices tested. The co-cultures made of amylolytic and probiotic bacteria strains are suitable to reduce the fermentation time of a soy milk/rice matrix and to obtain a final product with pH and viscosity similar to yoghurt.     

Evaluation of Pediococcus pentosaceus strains isolated from Idly batter for probiotic properties in vitro

Idly batter is a traditional fermented food in South India and distribution of lactic acid bacteria (LAB) varies with season. Six bacteriocinogenic Pediococcus pentosaceus strains isolated from Idly batter were evaluated for probiotic properties in vitro. These isolates exhibited effective inhibition against various intestinal Gram positive and negative pathogens like Escherichia coli. These isolates especially, VJ13, VJ31 and VJ49 could tolerate 0.3% bile salt, gastric and intestinal conditions. All isolates exhibited strong auto-aggregation and co-aggregation (against Listeria monocytogenes and E. coli) with higher hydrophobicity and thus they could eliminate pathogen and prevent its adhesion to intestinal cells. All isolates except VJ41 assimilate cholesterol effectively, suggesting that they can lower the cholesterol, and because they produce β-galactosidase they can be used for lactose intolerance. Heat-killed P. pentosaceus VJ13 also assimilated cholesterol (19%) but not as effectively as live strains (73%). Among these isolates, VJ13 and VJ49, which exhibited efficient probiotic property, were evaluated for in vitro adhesion with Caco-2 cells. These isolates adhere more effectively with Caco-2 cells than the intestinal pathogen L. monocytogenes and prevent the pathogen adhesion to intestinal cells. Among these isolates, VJ13 has beneficial probiotic properties which can be exploited for formulation of functional foods.     

Agar films containing green tea extract and probiotic bacteria for extending fish shelf-life

A bioactive film composed of agar, incorporating green tea extract and probiotic strains (Lactobacillus paracasei L26 and Bifidobacterium lactis B94) was applied on hake fillets in order to evaluate the effect of the films during 15 days of storage. Hake was previously inoculated with Shewanella putrefaciens and Photobacterium phosphoreum (10³–10⁴ CFU/g) to simulate a spoilage process. The green tea and/or probiotic film provoked a reduction, particularly of H₂S-producing bacteria counts and total viable bacteria throughout the storage period. The probiotic strains added to the film could pass to the fish producing an increment of lactic acid bacterial counts, even in the presence of green tea extract. The effect of the films also caused a decrease in the indexes of fish quality (total volatile basic nitrogen (TVB-N), trimethylamine nitrogen (TMA-N) and pH). The total viable counts, H₂S-producing microorganisms and TVB-N were maintained below the limits of acceptability during 15 days for the fillet covered with the green tea + probiotic film, compared to the rest of the samples. Films with green tea and probiotic were able to extend shelf-life of hake at least for a week and increase the beneficial lactic acid bacteria in fish.     

In vitro evaluation of the suitability potential probiotic of lactobacilli isolates from the gastrointestinal tract of chicken

Probiotic lactobacilli could be used to decrease the colonization of pathogenic bacteria in chicken and therefore decrease the risk of foodborne illness to consumers. The present study was conducted to select appropriate microbial strains for the development of potential probiotic. In experiment 1, 18 strains of lactobacilli isolated from the gastrointestinal tract of chicken were evaluated. The strains were demonstrated for their lactic acid, hydrogen peroxide, and exopolysaccharide productions. For experiment 2, the strains were tested for their acid, bile, antimicrobial activity, and antibiotic resistance levels. Among them, Lactobacillus delbrueckii ssp. delbrueckii BAZ32, Lactobacillus acidophilus BAZ29, BAZ36, BAZ43, and BAZ63 which produced high EPS were selected to aggregation ability. It is concluded that L. delbrueckii ssp. delbrueckii BAZ32, L. acidophilus BAZ29 confer high tolerance to acid, bile, antibiotic resistance, high antimicrobial activity, aggregation ability, and EPS production. These strains may be functional feed additives as potential probiotic in chicken.     

Selection of potential probiotic lactic acid bacteria from fermented olives by in vitro tests

The present study aims to evaluate the probiotic potential of lactic acid bacteria (LAB) isolated from naturally fermented olives and select candidates to be used as probiotic starters for the improvement of the traditional fermentation process and the production of newly added value functional foods. Seventy one (71) lactic acid bacterial strains (17 Leuconostoc mesenteroides, 1 Ln. pseudomesenteroides, 13 Lactobacillus plantarum, 37 Lb. pentosus, 1 Lb. paraplantarum, and 2 Lb. paracasei subsp. paracasei) isolated from table olives were screened for their probiotic potential. Lb. rhamnosus GG and Lb. casei Shirota were used as reference strains. The in vitro tests included survival in simulated gastrointestinal tract conditions, antimicrobial activity (against Listeria monocytogenes, Salmonella Enteritidis, Escherichia coli O157:H7), Caco-2 surface adhesion, resistance to 9 antibiotics and haemolytic activity. Three (3) Lb. pentosus, 4 Lb. plantarum and 2 Lb. paracasei subsp. paracasei strains demonstrated the highest final population (>8 log cfu/ml) after 3 h of exposure at low pH. The majority of the tested strains were resistant to bile salts even after 4 h of exposure, while 5 Lb. plantarum and 7 Lb. pentosus strains exhibited partial bile salt hydrolase activity. None of the strains inhibited the growth of the pathogens tested. Variable efficiency to adhere to Caco-2 cells was observed. This was the same regarding strains' susceptibility towards different antibiotics. None of the strains exhibited β-haemolytic activity. As a whole, 4 strains of Lb. pentosus, 3 strains of Lb. plantarum and 2 strains of Lb. paracasei subsp. paracasei were found to possess desirable in vitro probiotic properties similar to or even better than the reference probiotic strains Lb. casei Shirota and Lb. rhamnosus GG. These strains are good candidates for further investigation both with in vivo studies to elucidate their potential health benefits and in olive fermentation processes to assess their technological performance as novel probiotic starters.     

Identification and functional traits of lactic acid bacteria isolated from Ciauscolo salami produced in Central Italy

Lactic acid bacteria (LAB) from Ciauscolo salami produced in Marche Region of Central Italy, and LAB strains belonging to our laboratory collection were examined for their capability to survive at low pH and bile, to adhere to Caco-2 cells, and for antibiotic resistance. LAB from Ciauscolo were identified by ARDRA and RAPD-PCR. Our study showed that all LAB strains had good adaptation to gastric juice and moderate tolerance to bile. The adhesiveness was variable among strains but significantly lower in LAB from food. Antibiotic resistance was broadly spread among food strains, with level of resistance exceeding 15% for all the antibiotics tested. The resistance determinants erm(B) and tet(M) were found in nine strains of food origin (21.4%) while tet(L) in one strain of our collection (5%). Our work suggests that fermented foods are valuable sources of bacterial strains with functional traits of intestinal lactobacilli. These bacteria may be further studied for their use in probiotic applications.     

In vitro assessment of the gastrointestinal transit tolerance of taxonomic reference strains from human origin and probiotic product isolates of Bifidobacterium

Next to health promoting effects, the functional aspect of probiotic strains also involves their capacity to reach the colon as viable metabolically active cells. The present study aimed to assess the potential of 24 probiotic product isolates and 42 human reference strains of Bifidobacterium to survive gastrointestinal transit under in vitro conditions. The survival capacity of exponential and stationary phase cultures upon exposure to gastric and small intestinal juices was determined using a recently developed microplate-based assay in combination with the LIVE/DEAD BacLight Bacterial Viability kit. All 66 strains tested displayed a considerable loss in viability during exposure to an acidic pepsin containing solution (pH 2.0). Among the 10 taxa tested, cultures of B. animalis ssp. lactis appeared to be most capable to survive gastric transit. Although to a lesser extent, the presence of bile salts also affected the viability of most of the strains tested. Except for 3 strains, all 66 strains showed bile salt hydrolase activity using an agar-based assay. In contrast, the bifidobacterial strains used in this study appeared to possess a natural ability to survive the presence of pancreatin (pH 8.0). Although the effect was not significant, a slightly enhanced tolerance to gastrointestinal transit was observed when cells were in the stationary phase, especially when exposed to acid, compared with cells being in the exponential phase. Survival in the gastrointestinal tract appeared to be largely strain-dependent and hence implies that different strains will likely display a different behavior in functionality. The assay used in this study allows an initial assessment of strains for use as probiotic cultures prior to selecting potential candidate strains for further investigation in vivo.     

Probiotic properties of novel probiotic Levilactobacillus brevis KU15147 isolated from radish kimchi and its antioxidant and immune-enhancing activities

This study was conducted to evaluate the probiotic properties and antioxidant activities of lactic acid bacteria strains including Levilactobacillus brevis KU15147 isolated from kimchi to determine their potential as a probiotic. The tolerance of all strains to gastric acid and bile salts was more than 90%. The strains did not produce a β-glucuronidase and survived following treatment with gentamicin, kanamycin, streptomycin, and ciprofloxacin. L. brevis KU15147 showed greater adhesion activity to HT-29 cells (6.38%) and its antioxidant activities were higher than those of other tested strains, showing values of 38.56%, 22%, and 23.82% in DPPH, ABTS, and β-carotene bleaching assays, respectively. Additionally, the relative expression intensities of induced nitric oxide synthase and tumor necrosis factor-α of L. brevis KU15147 were greater than those of other strains, suggesting that this strain can be applied in the health food or pharmaceutical industry as a novel probiotic strain.     

Characterisation of the exopolysaccharide (EPS)-producing Lactobacillus paraplantarum BGCG11 and its non-EPS producing derivative strains as potential probiotics

Traditional fermented foods are the best source for the isolation of strains with specific traits to act as functional starters and to keep the biodiversity of the culture collections. Besides, these strains could be used in the formulation of foods claimed to promote health benefits, i.e. those containing probiotic microorganisms. For the rational selection of strains acting as probiotics, several in vitro tests have been proposed. In the current study, we have characterized the probiotic potential of the strain Lactobacillus paraplantarum BGCG11, isolated from a Serbian soft, white, homemade cheese, which is able to produce a "ropy" exopolysaccharide (EPS). Three novobiocin derivative strains, which have lost the ropy phenotype, were characterized as well in order to determine the putative role of the EPS in the probiotic potential. Under chemically gastrointestinal conditions, all strains were able to survive around 1-2% (10(6)-10(7)cfu/ml cultivable bacteria) only when they were included in a food matrix (1% skimmed milk). The strains were more resistant to acid conditions than to bile salts and gastric or pancreatic enzymes, which could be due to a pre-adaptation of the parental strain to acidic conditions in the cheese habitat. The ropy EPS did not improve the survival of the producing strain. On the contrary, the presence of an EPS layer surrounding the strain BGCG11 hindered its adhesion to the three epithelial intestinal cell lines tested, since the adhesion of the three non-ropy derivatives was higher than the parental one and also than that of the reference strain Lactobacillus rhamnosus GG. Aiming to propose a potential target application of these strains as probiotics, the cytokine production of peripheral blood mononuclear cells (PBMC) was analyzed. The EPS-producing L. paraplantarum BGCG11 strain showed an anti-inflammatory or immunosuppressor profile whereas the non-ropy derivative strains induced higher pro-inflammatory response. In addition, when PBMC were stimulated with increasing concentrations of the purified ropy EPS (1, 10 and 100μg/ml) the cytokine profile was similar to that obtained with the EPS-producing lactobacilli, therefore pointing to a putative role of this biopolymer in its immune response.