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Clinical studies & probiotics

CLINICAL STUDIES

EFFECT OF BIFIDOBACTERIUM BREVE ON THE INTESTINAL MICROBIOTA OF COELIAC CHILDREN ON A GLUTEN FREE DIET: A PILOT STUDY
ANDREA QUAGLIARIELLO 1, †, IRENE ALOISIO 2, †, NICOLE BOZZI CIONCI 2, DONATA LUISELLI 1, GIUSEPPE D’AURIA 3, LLÚCIA MARTINEZ-PRIEGO 3, DAVID PÉREZ-VILLARROYA 3, TOMAŽ LANGERHOLC 4, MAŠA PRIMEC 4, DUŠANKA MIˇCETI´C-TURK 5 AND DIANA DI GIOIA 2
ABSTRACT:
Abstract: Coeliac disease (CD) is associated with alterations of the intestinal microbiota. Although several Bifidobacterium strains showed anti-inflammatory activity and prevention of toxic gliadin peptides generation in vitro, few data are available on their efficacy when administered to CD subjects. This study evaluated the effect of administration for three months of a food supplement based on two Bifidobacterium breve strains (B632 and BR03) to restore the gut microbial balance in coeliac children on a gluten free diet (GFD). Microbial DNA was extracted from faeces of 40 coeliac children before and after probiotic or placebo administration and 16 healthy children (Control group). Sequencing of the amplified V3-V4 hypervariable region of 16S rRNA gene as well as qPCR of Bidobacterium spp., Lactobacillus spp., Bacteroides fragilis group Clostridium sensu stricto and enterobacteria were performed. The comparison between CD subjects and Control group revealed an alteration in the intestinal microbial composition of coeliacs mainly characterized by a reduction of the Firmicutes/Bacteroidetes ratio, of Actinobacteria and Euryarchaeota. Regarding the effects of the probiotic, an increase of Actinobacteria was found as well as a re-establishment of the physiological Firmicutes/Bacteroidetes ratio. Therefore, a three-month administration of B. breve strains helps in restoring the healthy percentage of main microbial components.
Probiotic Streptococcus thermophilus FP4 and Bifidobacterium breve BR03 Supplementation Attenuates Performance and Range-of-Motion Decrements Following Muscle Damaging Exercise
RALF JÄGER 1, MARTIN PURPURA 1, JASON D. STONE 2, STEPHANIE M. TURNER 2, ANTHONY J. ANZALONE 2, MICAH J. EIMERBRINK 2, MARCO PANE 3, ANGELA AMORUSO 3, DAVID S. ROWLANDS 4 AND JONATHAN M. OLIVER 2,
ABSTRACT:
Probiotics have immunomodulatory effects. However, little is known about the potential benefit of probiotics on the inflammation subsequent to strenuous exercise. In a double-blind, randomized, placebo controlled, crossover design separated by a 21-day washout, 15 healthy resistance-trained men ingested an encapsulated probiotic Streptococcus (S.) thermophilus FP4 and Bifidobacterium (B.) breve BR03 at 5 bn live cells (AFU) concentration each, or a placebo, daily for 3 weeks prior to muscle-damaging exercise (ClinicalTrials.gov NCT02520583). Isometric strength, muscle soreness, range of motion and girth, and blood interleukin-6 (IL-6) and creatine kinase (CK) concentrations were measured from pre- to 72 h post-exercise. Statistical analysis was via mixed models and magnitude-based inference to the standardized difference. Probiotic supplementation resulted in an overall decrease in circulating IL-6, which was sustained to 48 h post-exercise. In addition, probiotic supplementation likely enhanced isometric average peak torque production at 24 to 72 h into the recovery period following exercise (probiotic–placebo point effect 90% CI: 24 h, 11%  7%; 48 h, 12%  18%; 72 h, 8%  8%). Probiotics also likely moderately increased resting arm angle at 24 h (2.4%  2.0%) and 48 h (1.9%  1.9%) following exercise, but effects on soreness and flexed arm angle and CK were unclear. These data suggest that dietary supplementation with probiotic strains S. thermophilus FP4 and B. breve BR03 attenuates performance decrements and muscle tension in the days following muscle-damaging exercise.
Interaction between probiotics and human immune cells. The prospective anti-inflammatory activity of Bifidobacterium breve BR03.
STEFANIA NICOLA 1*, LUCA MOGNA 1, SERENA ALLESINA 1, MICHELA BARBA 1, FRANCESCA DEIDDA 1, PAOLA LORENZINI 1, ELISABETTA RAITERI 1, GIAN P. STROZZI 2, GIOVANNI MOGNA 1,2
ABSTRACT:
The human intestinal microflora may be considered as a post-natally acquired organ composed of a large diversity of bacteria with different functions on human health. Probiotics are live microorganisms which, when administered in adequate amounts, confer a health benefit on the host. Probiotics are widely used to rebalance microbiota composition, thus improving gut functionality and immune system responses. However, only few data on the specific immunologic pathways are currently available. This study showed that Bifidobacterium breve BR03 (DSM 16604) is able to induce a relevant increase of helper T lymphocytes and the secretion of Th2 cytokines (IL-10 and IL-4). This strain demonstrates a strong anti-inflammatory property through the shift of Th1/Th2 balance towards a Th2 response and could therefore be beneficial in irritable bowel diseases.
Assessment of the in vitro inhibitory activity of specific probiotic bacteria against different Escherichia coli strains.
Mogna L 1, Del Piano M, Deidda F, Nicola S, Soattini L, Debiaggi R, Sforza F, Strozzi G, Mogna G
ABSTRACT:
BACKGROUND:
Lactobacilli and bifidobacteria are often associated with health-promoting effects. These live microorganisms, defined as probiotics, are commonly consumed as part of fermented foods, such as yoghurt and fermented milks, or as dietary supplements. Escherichia coli is a gram-negative, rod-shaped bacterium commonly found in the lower intestine of warm-blooded organisms. As a part of the normal gut microbiota, this microorganism colonizes the gastrointestinal tract of animals and humans within a few hours after birth. All E. coli strains can produce a wide variety of biogenic amines responsible for potentially harmful systemic intoxications. Enterohemorrhagic E. coli serotype O157:H7 is a pathotype of diarrhoeagenic strains with a large virulence plasmid pO157 able to produce 1 or more Shiga toxins.
METHODS:
The overall aim of this study was to determine the inhibitory effects of different strains of probiotics on E. coli serotypes, including E. coli O157:H7 (CQ9485). In particular, the antagonistic activity of 4 Bifidobacterium strains (Probiotical SpA, Italy) and 16 lactic acid bacteria, more specifically 14 Lactobacillus spp. and 2 Streptococcus spp., was assessed against selected E. coli biotypes (ATCC 8739, ATCC 10536, ATCC 35218, and ATCC 25922). The diarrhoeagenic serotype O157:H7 was also tested. RESULTS: The experimental data collected demonstrated an in vitro significant inhibitory effect of 6 Lactobacillus strains, namely L. rhamnosus LR04, L. rhamnosus LR06, L. plantarum LP01, L. plantarum LP02, L. pentosus LPS01, and L. delbrueckii subsp. delbrueckii LDD01, and 2 Bifidobacterium strains, B. breve BR03 and B. breve B632. The inhibiting extent was slightly different among these strains, with L. delbrueckii subsp. delbrueckii LDD01 showing the highest activity on E. coli O157:H7.
CONCLUSIONS:
Most of the probiotics studied are able to antagonize the growth of the 5 strains of E. coli tested, including the O157:H7 biotype, well known for their characteristic to produce a wide variety of biogenic amines considered responsible for dangerous systemic intoxications.
Quantification of folic acid in human feces after administration of Bifidobacterium probiotic strains.
Strozzi GP 1, Mogna L.
ABSTRACT:
BACKGROUND:
Folic acid, or vitamin B9, is involved in appropriate regulation of DNA replication, synthesis of purines and deoxythymidine (dTMP), conversion of homocysteine to methionine, histidine catabolism, and correct differentiation of the neural tube during fetal organogenesis. Folic acid from food sources is almost completely absorbed in the small intestine, mostly in the jejunum, and does not reach the large intestine. The administration of probiotic strains able to synthesize folates de novo and release them in the extracellular space may provide an additional, constant endogenous source of this important vitamin in the intestinal lumen of humans.
METHODS:
A pilot study involving 23 healthy volunteers was conducted to evaluate the ability of 3 probiotic strains, Bifidobacterium adolescentis DSM 18350, B. adolescentis DSM 18352, and Bifidobacterium pseudocatenulatum DSM 18353, to produce folates in the human intestine. Volunteers were randomly assigned to 1 of 3 groups for treatment with a specific probiotic strain (5 x 10(9) colony forming units/d). Strain effectiveness was evaluated by determination of the folate concentration in feces evacuated within 48 hours before and after administration of the probiotics. Quantification of microorganisms belonging to the genus Bifidobacterium was performed in parallel to folate analysis.
RESULTS:
Ingestion of these probiotic strains resulted in a significant increase of folic acid concentration in human feces in all treated groups. Analysis of the fecal Bifidobacteria confirmed the potential of all strains, especially B. adolescentis DSM 18352, to colonize the intestinal environment.
CONCLUSIONS:
The demonstrated ability of the probiotic microorganisms B. adolescentis DSM 18350, B. adolescentis DSM 18352, and B. pseudocatenulatum DSM 18353 to synthesize and secrete folates in the human intestinal environment may provide a complementary endogenous source of such molecules, which is especially useful for the homeostasis of mucosal enterocytes of the colon and, unlike oral administration of the vitamin, ensures its constant bioavailability.
Can Lactobacillus fermentum LF10 and Lactobacillus acidophilus LA02 in a slow-release vaginal product be useful for prevention of recurrent vulvovaginal candidiasis?: A clinical study.
Murina F 1, Graziottin A, Vicariotto F, De Seta F.
ABSTRACT:
BACKGROUND:
To assess the effectiveness of the association of 2 specific strains, Lactobacillus fermentum LF10 (DSM 19187) and Lactobacillus acidophilus LA02 (DSM 21717), specifically formulated in slow-release effervescent tablets, in patients with recurrent vulvovaginal candidiasis.
STUDY DESIGN:
The study was a clinical trial of 58 women diagnosed with recurrent VVC (≥4 culture-confirmed episodes in a 12-mo period). All patients were given 200 mg of fluconazole orally as an induction dose for 3 alternate days during the first treatment week. Afterward, the patients were given a new product formulated in slow-release vaginal tablets containing at least 0.4 billion live cells of each of lactobacillus L. fermentum LF10 and L. acidophilus LA02 (first phase of the prophylactic period), on alternate days for 10 consecutive nights. Patients who were still free of symptoms were given 1 vaginal tablet every week for the next 10 weeks (second phase of the prophylactic period). Patients asymptomatic after the total duration of the observation phase (7 mo) were considered as responders.
RESULTS:
During the second 10-week prophylactic phase, 49 of 57 (86.0%) patients remained free of clinical recurrence, whereas symptomatic VVC occurred in 8 patients (14.0%). During the 7-month follow-up, 42 patients of 49 (85.7%) were symptom free at the end of the protocol, whereas clinical recurrences occurred in 7 women (14.3%). Overall, 42 of 58 women enrolled in the study (72.4%) experienced no clinical recurrence throughout the 7-month observation phase (responders).
CONCLUSIONS:
This study strengthens the evidence supporting the use of specific lactobacilli with well-demonstrated activities associated with the creation and maintenance of a vaginal biofilm that hinders the persistence of an infection caused by Candida.
Effectiveness of the association of 2 probiotic strains formulated in a slow release vaginal product, in women affected by vulvovaginal candidiasis: a pilot study.
Vicariotto F 1, Del Piano M, Mogna L, Mogna G.
ABSTRACT:
BACKGROUND:
Vulvovaginal candidiasis (VVC) is the second most common cause of vaginitis after bacterial vaginosis, and it is diagnosed in up to 40% of women with vaginal complaints in the primary care setting. Among Candida spp., Candida albicans is the most common infectious agent. The treatment of choice for uncomplicated VVC is achieved with single-dose or short-course therapy in over 90% of cases. Several topical and oral drugs are available, without evidence for superiority of any agent or route of administration. In any case, most classic treatments are unable to significantly offer a protection against possible recurrences. In recent years, probiotics are emerging as a new strategy to counteract VVC. In fact, they are well known for their ability to lower intravaginal pH, thus establishing a barrier effect against many types of yeasts. Some strains are also able to exert additional and more focused antagonistic activities mediated by specific molecules such as hydrogen peroxide and bacteriocins. For example, Lactobacillus fermentum LF5 (CNCM I-789) was successfully tested in 4 human trials involving a total of 340 women reporting VVC at enrollment. In any case, the way used to deliver probiotics to the vaginal environment represents a crucial point. The aim of this work was to first select 1 or more probiotic strains in vitro with an antagonistic activity on Candida yeasts and then to perform an in vivo human pilot study using an association of the most promising and active bacteria.
METHODS:
For this purpose, 2 probiotic strains Probiotical S.p.A (Italy) were selected based on their strong in vitro inhibition activity toward 4 particular Candida species, namely C. albicans, Candida glabrata, Candida parapsilosis, and Candida krusei and subsequently tested in a human intervention pilot trial involving 30 women with VVC. The probiotics used, L. fermentum LF10 (DSM 19187) and Lactobacillus acidophilus LA02 (DSM 21717), were administered by means of slow release effervescent vaginal tablets (ActiCand 30 product). The main endpoint was the assessment of the establishment and maintenance of a barrier effect against Candida yeasts in women suffering from VVC. Thirty female subjects who were diagnosed with VVC by both microscopic examination and yeast culture were enrolled in the study and directed to apply a vaginal tablet once a day for 7 consecutive nights, followed by 1 tablet every 3 nights for a further 3-week application (acute phase) and, finally, 1 tablet per week to maintain a long-term vaginal colonization against possible recurrences. A medical examination of each patient was performed at enrollment (d₀), at the end of the first 4 weeks of treatment (d₂₈), and at the end of the second month of relapse prevention (d₅₆). The visual and microscopic examination was always accompanied by microbiological analyses of vaginal swabs to assess the presence of Candida. A statistical comparison was made between d₂₈, or d₅₆, and d0, and between d₅₆ and d₂₈ to quantify the efficacy against possible recurrences.
RESULTS:
The administration of the product ActiCand 30 was able to significantly solve Candida yeast symptoms after 28 days in 26 patients out of 30 (corresponding to 86.6%, P<0.001). At the end of the second month, recurrences were recorded, albeit not particularly serious, in only 3 out of 26 patients (11.5%, P=0.083) who were found to have fully healed at the end of the first month of treatment. This is a further confirmation of the long-term barrier effect exerted by the product.
CONCLUSIONS:
VVC has a very high incidence as 70% to 75% of women report at least 1 episode during the life. Many treatments are currently available but, despite a relatively high effectiveness in the relief of symptoms typically associated with acute infections, they are generally unable to offer a long-term protective barrier against possible recurrences. This study demonstrated the ability of ActiCand 30 to not only solve Candida infections in a very high percentage of women, but also to exert a long-term physiological defense due to the colonization of vaginal microbiota and adhesion of the mucosa to the epithelial cells. The special formulation of ActiCand 30, consisting of slow release effervescent vaginal tablets, is able to mediate 2 types of barrier effects, the first represented by the formation of an anaerobic environment due to the release of CO₂ and the second guaranteed by the colonization and adhesion to the vaginal epithelium of the 2 probiotics L. fermentum LF10 and L. acidophilus LA02.
Effectiveness of the two microorganisms Lactobacillus fermentum LF15 and Lactobacillus plantarum LP01, formulated in slow-release vaginal tablets, in women affected by bacterial vaginosis: a pilot study.
Vicariotto F 1, Mogna L, Del Piano M.
ABSTRACT:
BACKGROUND:
Bacterial vaginosis (BV) is the most common reason for abnormal vaginal discharge in reproductive-age women and one of its most important causative agents is the gram-variable bacterium Gardnerella vaginalis. BV is not accompanied by significant local inflammation, whereas the "fishy odor" test is always positive. In contrast, aerobic vaginitis (AV) is predominantly associated with Escherichia coli, but Streptococcus agalactiae and Staphylococcus aureus are also involved. Standard treatment of BV consists of oral or intravaginal antibiotics, although these are unable to spontaneously restore normal flora characterized by a high concentration of lactobacilli. The main limitation is the inability to offer a long-term defensive barrier, thus facilitating relapses and recurrences. This study was undertaken firstly to assess the ability of selected lactobacilli to in vitro antagonize G. vaginalis to determine an association with a strain able to inhibit E. coli, thus identifying a possible use in AV. The second step of the study was to conduct a human pilot trial in women affected by BV using an association of the most promising and active bacteria.
MATERIALS AND METHODS:
For this purpose, neutralized supernatants of individual lactobacilli were tested at percentages ranging from 0.5% to 4% to determine their ability to hinder the growth of G. vaginalis American Type Culture Collection 10231. The bacterium that was able to exert the strongest inhibition was subsequently tested with Lactobacillus plantarum LP01 in a human intervention, placebo-controlled, pilot trial involving 34 female subjects (aged between 18 and 50, mean 34.7±8.9, no menopausal women) diagnosed with BV. The 2 microorganisms Lactobacillus fermentum LF15 (DSM 26955) and L. plantarum LP01 (LMG P-21021) were delivered to the vagina by means of slow-release vaginal tablets, also containing 50 mg of tara gum. The amount of each strain was 400 million live cells per dose. The women were instructed to apply a vaginal tablet once a day for 7 consecutive nights, followed by 1 tablet every 3 nights for a further 3-week application (acute phase) and, finally, 1 tablet per week to maintain a long-term vaginal colonization against possible recurrences. A clinical examination was performed and the Nugent score was quantified for each patient at enrollment (d0), after 28 days (d28), and at the end of the second month of relapse prevention (d56). A statistical comparison was made between d28, or d56, and d0, and between d56 and d28 to quantify the efficacy against possible recurrences.
RESULTS:
L. fermentum LF15 showed the strongest in vitro inhibitory activity towards G. vaginalis American Type Culture Collection (ATCC) 10231 after both 24 and 48 hours. In the human trial, the 2 lactobacilli selected, namely L. fermentum LF15 and L. plantarum LP01, significantly reduced the Nugent score below the threshold of 7 after 28 days in 22 patients of 24 in the active group (91.7%, P<0.001). Eight women (33.3%) recorded a Nugent score between 4 and 6, evidence of an intermediate situation, whereas the remaining 14 (58.3%) showed a score <4, therefore suggesting the restoration of physiological vaginal microbiota. At the end of the second month, only 4 women registered a Nugent score >7, definable as BV (16.7%, P=0.065 compared with d28). In the placebo group, no significant differences were recorded at any time.
CONCLUSIONS:
BV, also known as vaginal bacteriosis is the most common cause of vaginal infection in women of childbearing age. Furthermore, BV is often asymptomatic as about 50% of women with this condition have no symptoms at all and the prevalence rate in apparently healthy women is around 10%. This study suggests the ability of the 2 strains L. fermentum LF15 and L. plantarum LP01 to counteract acute Gardnerella infections effectively and significantly improve the related uncomfortable symptoms in a very high percentage of women. This could be partially attributed to the presence of tara gum, which is able to create a mechanical barrier against Gardnerella on the surface of vaginal mucosa as a primary mechanism. Furthermore, long-term physiological protection seems to be established, thanks to the integration of the 2 lactobacilli into the vaginal microbiota and to their adhesion to the epithelial cells of the mucosa. In the light of the additional in vitro inhibitory activity against E. coli, their prospective use in AV could also prove interesting.
Capability of the two microorganisms Bifidobacterium breve B632 and Bifidobacterium breve BR03 to colonize the intestinal microbiota of children.
Mogna L 1, Del Piano M, Mogna G.
ABSTRACT:
BACKGROUND:
The total number of bacteria present in the gut microbiota of a newborn is consistently lower than the average found in adults, with the extent of this difference being directly related to body weight and age. It could be assumed that a lower number of viable probiotic cells is necessary to achieve significant gut colonization in infants and children. This study assessed the capability of Bifidobacterium breve B632 (DSM 24706) and Bifidobacterium breve BR03 (DSM 16604), 2 strains able to significantly inhibit some gram-negative bacteria in vitro, to integrate into the intestinal microbiota of children.
MATERIALS AND METHODS:
Ten healthy children aged an average of 5.7±2.6 were given an oily suspension containing B. breve B632 and B. breve BR03 for 21 consecutive days. The daily dose was 100 million live cells of each strain. Fecal specimens were collected and analyzed at the beginning (d0) and at the end of the study (d21). Total fecal bifidobacteria and coliforms have been quantified by microbiological plate counts.
RESULTS:
A significant increase in total fecal bifidobacteria (from 8.99 to 9.47 log10 CFU/g, P=0.042) and a parallel decrease in total coliforms (from 8.60 to 7.93 log10 CFU/g, P=0.048) was recorded after 21 days of supplementation.
CONCLUSIONS:
An oily suspension has proved an effective way of providing probiotics to children. A lower viable cells concentration was sufficient to mediate this effect in the light of the fact that the intestinal microbiota of children harbors a considerably smaller amount of total bacteria compared with adults. In addition to gut colonization in healthy children, B. breve B632 and B. breve BR03 were able to decrease total fecal coliforms, therefore supporting their potential specific use in colicky infants.
Effectiveness of an association of a cranberry dry extract, D-mannose, and the two microorganisms Lactobacillus plantarum LP01 and Lactobacillus paracasei LPC09 in women affected by cystitis: a pilot study.
Vicariotto F.
ABSTRACT:
BACKGROUND:
Urinary tract infections (UTIs) are the most common bacterial infection in women. Most UTIs are acute uncomplicated cystitis caused by Escherichia coli (86%). This study was undertaken to assess the effectiveness of an association of a cranberry dry extract, D-mannose, a gelling complex composed of the exopolysaccharides produced by Streptococcus thermophilus ST10 (DSM 25246) and tara gum, as well as the 2 microorganisms Lactobacillus plantarum LP01 (LMG P-21021) and Lactobacillus paracasei LPC09 (DSM 24243) in women affected by acute uncomplicated cystitis.
MATERIALS AND METHODS:
Thirty-three premenopausal, nonpregnant women diagnosed with acute uncomplicated cystitis were enrolled in a pilot prospective study and completed the treatment protocol. Subjects were instructed to take 2 doses per day during the first month, and then to continue with 1 sachet per day until the sixtieth day. Nitrites and leukocyte esterase on urine dipstick testing were used as indicators of cystitis, with analysis performed at enrollment, after 30 and 60 days, and after 1 month of follow-up. Typical UTI symptoms, namely dysuria, frequent voiding of small volumes, urinary urgency, suprapubic pain, and gross hematuria were scored 0 to 3 and evaluated at each visit.
RESULTS:
Positive results for the presence of nitrites and leukocyte esterase were found in 14 and 20 subjects after 30 days and in 9 and 14 women after 60 days, respectively <P<0.001). At the end of the follow-up period, positive results for nitrites and leukocyte esterase were recorded in only 4 and 3 of 24 and 19 subjects (16.7%, P=0.103; 15.8%, P=0.325, respectively), with negative results after 60 days. Typical symptoms of cystitis, specifically dysuria, frequent voiding, urgency, and suprapubic pain were significantly improved as well. No significant differences were recorded in the incidence and severity of hematuria at any visit.
CONCLUSION:
The long-term ability of an association of cranberry, D-mannose, an innovative gelling complex, and the 2 microorganisms tested to significantly improve the uncomfortable symptoms reported by women with acute cystitis has been suggested.
Bifidobacteria supplementation: effects on plasma lipid profiles in dyslipidemic children.
Guardamagna O1, Amaretti A2, Puddu PE3, Raimondi S2, Abello F4, Cagliero P4, Rossi M2.
ABSTRACT:
OBJECTIVE:
Preclinical investigations support the use of probiotics in the treatment of hypercholesterolemia, but clinical evidence is often contrasting. The aim of this study was to evaluate the effects of a probiotic formulation containing three Bifidobacterium strains on lipid profiles in children affected by primary dyslipidemia.
METHODS:
Thirty-eight children with dyslipidemia, ages 10.8 ± 2.1 y, were enrolled in a randomized, double-blind, placebo-controlled cross-over study. After a 4-wk diet run-in period, the children received probiotics (B. animalis subspecies lactis MB 2409, B. bifidum MB 109B, and B. longum subspecies longum BL04) or placebo for 3 mo. After 1 mo, wash-out treatments were switched. A strict dietary evaluation concerning satured fatty acids and cholesterol content, STEP I diet accordingly, was performed by a dietitian who examined the weekly dietary diary at each visit.
RESULTS:
Baseline lipid profile was (mean ± SD): total cholesterol (TC) 222.8 ± 23.2 mg/dL, high-density lipoprotein cholesterol (HDL-C) 55.8 ± 12.2 mg/dL, triglycerides (TG) 99.0 ± 61.7 mg/dL, and low-density lipoprotein cholesterol (LDL-C) 147.2 ± 21.9 mg/dL. After 3 mo of probiotic treatment, the lipid profile was: TC 211.9 ± 27.3 mg/dL, HDL-C 60.7 ± 14.2 mg/dL, TG 79.5 ± 34.5 mg/dL, and LDL-C 135.3 ± 24.2 mg/dL. Compared with placebo, probiotics reduced TC by 3.4% (P = 0.02) and LDL-C by 3.8% (P = 0.001). No significant dietary change occurred through the study and no relevant adverse effects were observed.
CONCLUSIONS:
Treatment with a Bifidobacterium probiotic formulation was well tolerated and useful in combination with to diet therapy. Children with dyslipidemia benefited from this approach, although the results need to be confirmed by larger controlled studies. Copyright © 2014 Elsevier Inc. All rights reserved.
Cultivable and pyrosequenced fecal microflora in centenarians and young subjects.
Drago L 1, Toscano M, Rodighiero V, De Vecchi E, Mogna G.
ABSTRACT:
GOAL:
The aim of this study was to characterize the composition of the intestinal microbiota in healthy centenarians in comparison with younger adults, considering both quantitative and qualitative aspects of gut community structure.
BACKGROUND:
The gut microbiota plays an essential role in human health. Toward seniority, its balance is affected by deep physiological changes. Long-lived people (age >90 y) have unusual features that differ from the younger elderly, so they should be considered separately when analyzing age-related features. However, they have been included in few studies and they have usually been grouped together with the younger elderly.
STUDY:
The gut microbiota of 14 centenarians and 10 younger adults was analyzed. Cultivable bacteria belonging to the following groups were enumerated: enterobacteriaceae, Enterococcus, Staphylococcus, Lactobacillus, Bifidobacterium, Clostridium, Bacteroides, and yeast. Lactobacilli and Bifidobacteria were further characterized at the species level by pyrosequencing.
RESULTS:
In centenarians, we observed a reduction in the quantity of enterobacteriaceae, bifidobacteria, and bacteroides and an increase in clostridia sensu stricto (P<0.05). The number of Lactobacillus and Bifidobacterium species isolated in centenarians and younger adults was similar. The composition of the Lactobacillus subpopulation was quite different between the groups. The presence of Bifidobacterium longum in the gut seems to be a particular feature in centenarians. It is interesting to note that only 1 strain of B. longum was isolated from each centenarian subject.
CONCLUSIONS:
The gut microbiota of centenarians has particular features that differ from both younger adults and the younger elderly. Further studies would help to understand whether the intestinal microbiota can influence life expectancy and whether the administration of probiotic bacteria could help to extend the longevity of human life.
Selenium and zinc internalized by Lactobacillus buchneri Lb26 (DSM 16341) and Bifidobacterium lactis Bb1 (DSM 17850): improved bioavailability using a new biological approach.
Mogna L 1, Nicola S, Pane M, Lorenzini P, Strozzi G, Mogna G.
ABSTRACT:
BACKGROUND:
Minerals, often referred to as micronutrients, are one of the 5 fundamental groups of nutrients needed to sustain life. Micronutrient malnutrition affects >50% of the worldwide population. In particular, zinc (Zn) deficiency is considered an emerging public health problem in India and in other developing countries. Selenium (Se) is another trace mineral essential for humans and animals. Dietary Se exists primarily as selenomethionine and selenocysteine. In addition, Se may be present in its inorganic form (selenite) in some vegetables. To increase the daily intake of these minerals, numerous food supplements containing different inorganic and organic forms of Zn or Se are commercially available. At any rate, it is quite well known that inorganic salts have a very low bioavailability. Organic salts, commonly based on gluconate, orotate, citrate, or other molecules, are characterized by a higher systemic effect. The innovative opportunity of using certain species of probiotics enriched with the 2 minerals could represent an interesting alternative to these preparations. Diet integration with bacteria able to internalize Zn and Se may embody a new application of probiotics.
METHODS:
To overcome the difficulties of in vivo animal or human trials, in this work a cell culture model using Caco-2 cells in bicameral chambers (Transwell system) was developed and validated to quantify the bioavailability of some commercial forms of Se and Zn compared with the organic forms accumulated intracellularly by Lactobacillus buchneri Lb26 (DSM 16341) and Bifidobacterium lactis Bb1 (DSM 17850), respectively.
RESULTS:
The experimental data collected demonstrated a significantly higher bioavailability of Se and Zn internalized by L. buchneri Lb26 (DSM 16341) and B. lactis Bb1 (DSM 17850), respectively, compared with the inorganic and even organic forms tested. In particular, the Se accumulated at the intracellular level by L. buchneri Lb26 proved to be 5.9, 9.4, and 65 times more absorbable than sodium selenite, seleno-L-methionine, and seleno-L-cysteine, respectively. In contrast, Zn internalized by B. lactis Bb1 showed an absorption that was >16 times higher by Caco-2 cells compared with zinc gluconate and a 31.5 times higher absorption compared with zinc sulfate. Most notably, Se and Zn internalized by the 2 probiotics studied are the only forms able to reach the Transwell basolateral compartment at a concentration higher than the concentration found in the apical compartment, therefore suggesting a considerably higher in vivo ability to be absorbed into the bloodstream. Both organic and inorganic forms of Se and Zn were predominantly found in the apical compartment, thus demonstrating their poor ability to diffuse into the cell and become bioavailable in all subcellular areas.
CONCLUSIONS:
The opportunity of delivering minerals in a highly bioavailable form by means of a probiotic bacterium has not been deeply investigated to date. This is the first study reporting quantitative data on the bioavailability and percentage of absorption of minerals internalized by specific probiotics. The most noticeable aspect is the significantly higher absorption of both probiotic Se and Zn compared with their organic forms, with particular reference to seleno-L-methionine, seleno-L-cysteine, and zinc gluconate.
Allergen free probiotics
Mogna G 1, Strozzi GP, Mogna L.
ABSTRACT:
Food sensitivities are constantly increasing in "westernized" countries and may pose serious health risks to sensitized individuals. Severe allergy episodes have also been reported after the intake of probiotic products containing milk protein residues, especially in children. The need for safe and effective probiotic strains and food supplements, which contain them, is now emerging clearly. The present work describes the way of achieving this aim by the avoidance of any kind of raw materials at risk, both in probiotic strain industrial manufacturing and finished product formulation. Allergen-free probiotics represent, without any doubt, an innovative and safe tool for human health.
A new chance of preventing winter diseases by the administration of synbiotic formulations.
Pregliasco F 1, Anselmi G, Fonte L, Giussani F, Schieppati S, Soletti L.
ABSTRACT:
BACKGROUND:
The efficacy of probiotics is currently well documented with regard to the improvement of gastrointestinal functions, whereas their potential role in the prevention of infectious respiratory diseases has not been sufficiently analyzed. PURPOSE OF THE STUDY AND METHODS: A 3-stage prospective, randomized, double blind, placebo-controlled study was carried out with several synbiotic preparations containing 3 to 5 strains of Lactobacillus plantarum, Lactobacillus rhamnosus, and Bifidobacterium lactis, lactoferrin and prebiotics such as either FOS (short-chain fructooligosaccharides) or GOS (galactooligosaccharides). The study was performed over 3 different winter seasons between 2003 and 2007, and was aimed at assessing the ability of the different preparations to improve intestinal functions and to increase the body’s defences against respiratory infections. In 2003/04 (stage 1; 237 healthy volunteers) an active formulation (A) containing 3 probiotic strains and FOS was used versus placebo; in 2005/06 (stage 2; 234 healthy volunteers) the same formulation versus a similar preparation enriched with lactoferrin (B), and versus placebo; in 2006/07 (stage 3; 250 healthy volunteers), 2 new synbiotic formulations each containing 5 probiotics and FOS (C) or GOS (D), respectively, versus placebo.
RESULTS:
In stage 1, bowel functions improved (P=0.004) in terms of reduced bloating and more regular intestinal motility. The length of acute respiratory infection episodes considered as a whole (-0.97 d; P=0.007) and upper respiratory tract infections (URTIs, -1.96 d; P=0.044) were significantly decreased in the synbiotic group. The severity of episodes recorded a statistically significant drop in both episodes considered as a whole (3.21 average score vs. 3.98 in the placebo group, P<0.001) and in URTI (2.56 vs. 3.82; P=0.004) and flu classes (3.80 vs. 4.67, P=0.001). In stage 2, improvement of bowel functions was statistically significant (P=0.005) in synbiotic preparation A. A statistically significant reduction in the number of respiratory tract infections episodes was noted with both the two active formulations (P=0.002 in group A and P=0.003 in group B). The duration of episodes considered as a whole (-1.12 d in one of the 2 active formulation groups; P=0.005), URTIs (-2.08 d in group A; P=0.036) and influenza-like illness episodes (-1.40 d in group A; P=0.049) was significantly decreased in the synbiotic group. A reduction trend in cold episodes was also recorded. The severity of episodes recorded a statistically significant drop in episodes considered as a whole (-0.73 in group A, P=0.003; -0.65 in group B, P=0.004) and in the case of flu (-1.25 in group A, P<0.001; -1.18 in group B, P<0.001). In stage 3, the improvement of bowel functions was confirmed for both active formulations (P<0.001). A significant decrease in the total length of respiratory episodes (-1.51 d; P<0.001 in the group C and -1.39 d; P<0.001 in the group D) and in the length of cough (-3.08 d; P<0.001 in group C; -2.83 d; P<0.001 in group D), cold (-1.02 d; P=0.019 in group C; -1.32 d; P=0.001 in group D) and flu episodes was reported. The severity of episodes recorded a statistically significant drop in regard to episodes considered as a whole, the cold and flu classes in both groups and for cough too in group C. The number of episodes also dropped considerably in terms of overall episodes, cold (group C) and flu.
CONCLUSIONS:
These results demonstrate that a regular, long-term intake of various synbiotics may improve health by reducing the incidence and severity of respiratory diseases during the cold season.
Characterization of Bifidobacterium spp. strains for the treatment of enteric disorders in newborns.
Aloisio I 1, Santini C, Biavati B, Dinelli G, Cencič A, Chingwaru W, Mogna L, Di Gioia D.
ABSTRACT:
Several studies support the use of probiotics for the treatment of minor gastrointestinal problems in infants. Positive effects on newborn colics have been evidenced after administration of Lactobacillus strains, whereas no studies have been reported regarding the use of bifidobacteria for this purpose. This work was therefore aimed at the characterization of Bifidobacterium strains capable of inhibiting the growth of pathogens typical of the infant gastrointestinal tract and of coliforms isolated from colic newborns. Among the 46 Bifidobacterium strains considered, 16 showed high antimicrobial activity against potential pathogens; these strains were further characterized from a taxonomic point of view, for the presence and transferability of antibiotic resistances, for citotoxic effects and adhesion to nontumorigenic gut epithelium cell lines. Moreover, their ability to stimulate gut health by increasing the metabolic activity and the immune response of epithelial cells was also studied. The examination of all these features allowed to identify three Bifidobacterium breve strains and a Bifidobacterium longum subsp. longum strain as potential probiotics for the treatments of enteric disorders in newborns such as infantile colics. A validation clinical trial involving the selected strains is being planned.
Cholesterol-lowering probiotics: in vitro selection and in vivo testing of bifidobacteria.
Bordoni A 1, Amaretti A, Leonardi A, Boschetti E, Danesi F, Matteuzzi D, Roncaglia L, Raimondi S, Rossi M.
ABSTRACT:
Thirty-four strains of bifidobacteria belonging to Bifidobacterium adolescentis, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, and Bifidobacterium pseu-docatenulatum were assayed in vitro for the ability to assimilate cholesterol and for bile salt hydrolase (BSH) against glycocholic and taurodeoxycholic acids (GCA and TDCA). Cholesterol assimilation was peculiar characteristic of two strains belonging to the species B. bifidum (B. bifidum MB 107 and B. bifidum MB 109), which removed 81 and 50 mg of cholesterol per gram of biomass, being the median of specific cholesterol absorption by bifidobacteria 19 mg/g. Significant differences in BSH activities were not established among bifidobacterial species. However, the screening resulted in the selection of promising strains able to efficiently deconjugate GCA and TDCA. No relationship was recognized between BSH phenotype and the extent of cholesterol assimilation. On the basis of cholesterol assimilation or BSHGCA and BSHTDCA activities, B. bifidum MB 109 (DSMZ 23731), B. breve MB 113 (DSMZ 23732), and B. animalis subsp. lactis MB 2409 (DSMZ 23733) were combined in a probiotic mixture to be fed to hypercholesterolemic rats. The administration of this probiotic formulation resulted in a significant reduction of total cholesterol and low-density cholesterol (LDL-C), whereas it did not affect high-density cholesterol (HDL-C) and HDL-C/LDL-C ratio.
THE USE OF PROBIOTICS IN THE TREATMENT OF CONSTIPATION IN THE ELDERLY
M. Del Piano *, F. Montino*, S. Carmagnola*, A. Anderloni*, M. Orsello*, E. Garello*, F. Sforza**, M. Ballare* * Gastroenterology Independent Operating Unit, Hospital “Maggiore della Carità”, Novara. ** Nursing Home “I Cedri”, Fara Novarese.
The use of probiotics in healthy volunteers with evacuation dis orders and hard stools: a double-blind, randomized, placebo-controlled study.
Del Piano M 1, Carmagnola S, Anderloni A, Andorno S, Ballarè M, Balzarini M, Montino F, Orsello M, Pagliarulo M, Sartori M, Tari R, Sforza F, Capurso L.
ABSTRACT:
BACKGROUND:
Evacuation disorders and hard stools are common in industrialized countries, affecting on average 12% to 17% of the adult healthy population at any age. Dietary supplementation with probiotic microorganisms may be useful in reducing the disorder.
METHODS:
We performed a double-blind, randomized, placebo-controlled study to evaluate the effectiveness of 2 different probiotic blends, either mixed Lactobacillus plantarum LP01 (LMG P-21021) and Bifidobacterium breve BR03 (DSM 16604) or Bifidobacterium animalis subspecies lactis BS01 (LMG P-21384), in the management of evacuation disorders and intestinal discomfort. In a period of 5 years (2003 to 2008), the study involved 300 healthy volunteers (151 males and 149 females; age 24 to 71 y) with evacuation disorders and hard stools. In particular, subjects were divided into 3 groups: 80 subjects in the group A received placebo, 110 subjects in the group B received mixed L. plantarum LP01 and B. breve BR03 (2.5 x 10 colony-forming units/d of each strain), and 110 subjects in the group C received B. animalis subsp. lactis BS01 (5 x 10 colony-forming units/d) for 30 days. At the beginning of the observational study, the healthy status of volunteers was evaluated by a complete, laboratory and ultrasound study of the abdomen. The physical examination was repeated after 15 and 30 days. In particular, the main troubles typically associated with evacuation disorders and hard stools as well as abdominal bloating were considered as parameters of interest. Exclusion criteria were items of gastrointestinal diseases and antibiotics intake.
RESULTS:
Subjects treated with the mixed probiotic strains L. plantarum LP01 and B. breve BR03 or B. animalis subsp. lactis BS01 reported a significant improvement in the number of weekly bowel movements and in the main troubles associated with evacuations, particularly consistency of feces and ease of expulsion. Discomfort items such as abdominal bloating and anal itching, burning, or pain also registered a relevant improvement in the active groups receiving probiotics.
CONCLUSIONS:
The intake of an effective amount of mixed L. plantarum LP01 and B. breve BR03 or B. animalis subsp. lactis BS01 for 30 days is able to significantly relieve the evacuation disorders and hard stools, thus providing a useful tool for the management of such condition, which is particularly widespread in industrialized countries at any age.
Correlation between chronic treatment with proton pump inhibitors and bacterial overgrowth in the stomach: any possible beneficial role for selected lactobacilli?
Del Piano M 1, Pagliarulo M, Tari R, Carmagnola S, Balzarini M, Lorenzini P, Pane M.
ABSTRACT:
BACKGROUND:
The inhibition of physiological gastric acid secretion induced by proton pump inhibitors (PPIs), the most widely used drugs in the world, may cause a significant bacterial overgrowth in the gastrointestinal tract as a side effect. This study was undertaken firstly to correlate PPI intake with concentration of specific bacterial groups in the stomach as well as possible Helicobacter pylori infection, and secondly to assess the efficacy of the 4 lactobacilli L. rhamnosus LR06 (DSM 21981), L. pentosus LPS01 (DSM 21980), L. plantarum LP01 (LMG P-21021), and L. delbrueckii subsp. delbrueckii LDD01 (DSM 22106) in the restoration of a physiological gastric barrier.
METHODS:
Total bacteria, sulphite-reducing bacteria (SRB), total coliforms, and total lactobacilli were quantified in samples of gastric juice from 29 subjects taking PPIs for at least 3 months compared with 36 control subjects. The presence of H. pylori was also assessed.The subjects treated with PPIs with a concentration of total bacteria in the gastric juice higher than 10(5) cells/mL were selected for an intervention study with the 4 lactobacilli L. rhamnosus LR06, L. pentosus LPS01, L. plantarum LP01, and L. delbrueckii subsp. delbrueckii LDD01. After 15 days of supplementation, the same bacterial groups were quantified to compare these values with the baseline.
RESULTS:
No significant correlation was found between the presence of H. pylori and PPI intake. The baseline quantification of bacterial groups (log10 CFU/mL of gastric juice, PPI group vs. control) showed: total bacteria 8.35 versus 3.95 (P<0.001); total coliforms 4.98 versus 2.35 (P<0.001); SRB 5.71 versus 2.28 (P=0.065); and total lactobacilli 3.85 versus 2.20 (P=0.005). After 15 days of treatment with the 4 lactobacilli, the quantification of bacterial groups gave the following results: total bacteria 7.91 versus 8.35 at time zero (P=0.002); total coliforms 4.21 versus 4.98 at time zero (P<0.001); SRB 4.94 versus 5.71 at baseline (P=0.060); and total lactobacilli 7.20 versus 3.85 at baseline (P=0.040).
CONCLUSIONS:
A significant impairment of intragastric acidity is sufficient to induce a relevant bacterial overgrowth, with particular reference to SRB and total coliforms. This fact can contribute to an increase in the risk of infections and intestinal diseases. It could be crucial to restore the physiological "gastric barrier." The 2-week supplementation with the 4 lactobacilli tested proved to be effective in significantly reducing total bacteria and coliforms in the gastric milieu in subjects chronically treated with PPIs. It is therefore possible to hopothesise a beneficial role for such lactobacilli in clinical practice.
Is microencapsulation the future of probiotic preparations? The increased efficacy of gastro-protected probiotics.
Del Piano M 1, Carmagnola S, Ballarè M, Sartori M, Orsello M, Balzarini M, Pagliarulo M, Tari R, Anderloni A, Strozzi GP, Mogna L, Sforza F, Capurso L.
ABSTRACT:
In a recent publication we assessed the kinetics of intestinal colonization by microencapsulated probiotic bacteria in comparison with the same strains given in an uncoated form. It's well known, in fact, that microencapsulation of probiotics with specific materials is able to confer a significant resistance to gastric juice, thus protecting the cells during the gastric and duodenal transit and enhancing the probiotic efficacy of any supplementation. In any case, this was the first study reporting the fecal amounts of probiotics administered in a coated, protected form compared with traditional, uncoated ones. Here we discuss additional in vitro data of resistance of the same bacteria to gastric juice, human bile and pancreatic secretion and correlate them with the results of in vivo gut colonization.
Probiotics and chronic diarrhoea in the elderly
A. Dezi*, S. Carmagnola**, M. Ballarè**, M. Orsello**, M. Del Piano**, L. Capurso* * Gastroenterology Department, San Filippo Neri Hospital, Rome; ** Gastroenterology Department, Maggiore della Carità Hospital, Novara
Diet and chronic constipation. Benefits of oral supplementation with symbiotic zir fos (Bifidobacterium longum W11 + FOS Actilight).
Amenta M 1, Cascio MT, Di Fiore P, Venturini I.
ABSTRACT:
BACKGROUND AND AIM:
Constipation is one of the most common disorders in Western countries and it is known that dietary factors such as a low fiber diet and low caloric intake are associated with this condition. Weight loss can disrupt the intestinal ecosystem resulting in intestinal dysbiosis that worsens constipation. The aim of this study was to evaluate the effects of treatment with symbiotic zir fos (Bifidobacterium longum W11 + FOS Actilight) on chronic constipation in patients undergoing a weight loss diet.
METHODS:
Our evaluation included the recording of age, gender, weight, height, BMI, physical activity, constipation, diet, therapy compliance and laxative supplies. A hypocaloric diet (1,200/1,400 cal.) was prescribed to all patients, and they were submitted to a physical activity program and received 1 bag of symbiotic zir fos per day for the entire duration of the study. Patients' follow-up was available for up to 60 days.
RESULTS:
Two hundred and ninety seven patients (79.4% women and 18.2% men, mean age 32.2) were included in the study. The mean baseline BMI was 33.4 +/- 5.6 (range 22.8-56.3 Kg). The improvement of constipation turned out to be associated to age (p < 0.01). Patients with a mean age of 35 +/- 12 showed an improvement of constipation. BMI values were not significantly different among the groups of patients with improved, worsened or unchanged constipation. No significative difference was observed among groups due to physical activity. At the follow-up, after 20 days from the beginning of the study, patients that assumed at least 17/20 of the zir fos bags showed a greater improvement of constipation (p < 0.01) than the remaining patients who assumed less than 17/20 of the zir fos bags or that didn't assumed any at all. Patients that assumed laxatives (at least once a week) showedn to be more frequently associated with a worsening of constipation (p < 0.001). Diet compliance does not seem to influence the course of constipation.
CONCLUSION:
Our data demonstrate the utility of symbiotics in improving constipation during hypocaloric diet in the treatment of obesity.
Effect of a symbiotic preparation on the clinical manifestations of irritable bowel syndrome, constipation-variant. Results of an open, uncontrolled multicenter study.
Colecchia A 1, Vestito A, La Rocca A, Pasqui F, Nikiforaki A, Festi D; Symbiotic Study Group.
ABSTRACT:
AIM:
Irritable bowel syndrome (IBS) is frequently associated with an imbalance in intestinal bacteria. To date, few studies have evaluated the efficacy and safety of probiotic administration in patients with constipation-variant IBS. A new agent recently available in clinical practice is a symbiotic consisting of a probiotic, Bifidobacterium longum W11, and the short chain oligosaccharide prebiotic Fos Actilight. The aim of this study was to evaluate the efficacy and safety of this symbiotic in patients with constipation-variant IBS.
METHODS:
A total of 636 patients (250 men, 386 women) diagnosed with constipation-type IBS according to the Roma II criteria were enrolled in 43 centers and received the symbiotic at a dose of 3 g/die for at least 36 days. A validated questionnaire investigating symptoms and stool frequency was administered before and after treatment.
RESULTS:
Based on patient responses to visual scale items, frequency increased significantly after treatment in the ''no symptom'' class from 3% to 26.7% for bloating and from 8.4% to 44.1% for abdominal pain (P<0.0001). In the more severe symptoms classes (moderate-severe), symptom frequency dropped significantly from 62.9% to 9.6% and from 38.8% to 4.1% for bloating and abdominal pain, respectively. Stool frequency significantly increased from 2.9+/-1.6 times/week to 4.1+/-1.6 times/ week.
CONCLUSION:
The study product can increase stool frequency in patients with constipation-variant IBS and reduce abdominal pain and bloating in those with moderate-severe symptoms.
Changing of fecal flora and clinical effect of L. salivarius LS01 in adults with atopic dermatitis.
Drago L 1, Toscano M, De Vecchi E, Piconi S, Iemoli E.
ABSTRACT:
GOAL:
To evaluate cytokine stimulation with 3 strains of Lactobacillus salivarius in vitro and to assess changes in intestinal microflora and clinical improvements in adults with atopic dermatitis (AD) induced by the strain showing the best immunomodulatory features.
BACKGROUND:
AD is a common skin disease in children and adults. It is characterized by chronic inflammation, eczema, and increasing intestinal permeability. Various studies have shown that patients with AD presented some modifications in the intestinal microbiota composition; as a result, intestinal microflora is thought to have a pivotal role in this disease.
METHODS:
Thirty-eight patients aged from 18 to 46 years with moderate/severe AD were recruited. Subjects were randomized in a double-blind placebo-controlled study to receive active treatment with L. salivarius LS01: probiotic (n=19) or placebo (n=19). Cytokine production was determined by means of specific quantitative enzyme-linked immunosorbent assays. Intestinal bacterial groups were quantified using conventional culture techniques, whereas L. salivarius LS01 was identified using polymerase chain reaction and pulse field gel electrophoresis.
RESULTS:
L. salivarius LS01 showed the best immunomodulatory features and it was chosen for the second phase of the study. AD subjects showed a reduction in their SCORAD score after probiotic treatment and a significant decrease in the staphylococci load compared with the placebo group. Moreover, L. salivarius LS01 showed the ability to reduce the production of Th2 cytokines, maintaining the production of Th1 cytokines stable.
CONCLUSION:
Treatment with the L. salivarius LS01 strain seems to positively modify clinical and immunologic status and dermatology life quality in a group of adults affected by moderate/severe AD, leading to a rebalancing of altered intestinal microbiota.
Preliminary results on clinical effects of probiotic Lactobacillus salivarius LS01 in children affected by atopic dermatitis.
Niccoli AA 1, Artesi AL, Candio F, Ceccarelli S, Cozzali R, Ferraro L, Fiumana D, Mencacci M, Morlupo M, Pazzelli P, Rossi L, Toscano M, Drago L.
ABSTRACT:
GOAL:
The goal of this study was to evaluate the clinical efficacy of an intake of Lactobacillus salivarius LS01 (DSM 22775) for the treatment of atopic dermatitis (AD) in children.
BACKGROUND:
AD is an inflammatory and pruritic chronic relapsing skin disorder with multifactorial etiopathology. Some evidence suggests that probiotics may improve AD by modulating the immune system and the composition of intestinal microbiota.
STUDY:
A total of 43 patients aged from 0 to 11 years were enrolled in the study (M/F ratio=1:1) and treated with the probiotic strain L. salivarius LS01. Clinical efficacy of probiotic treatment was assessed from baseline by changes in itch index and in the objective SCORAD/SCORAD index.
RESULTS:
Patients being given probiotic treatment showed a significant improvement in clinical parameters (SCORAD and itch values) from baseline. The reduction in SCORAD and itch index observed after 4 weeks of treatment also persisted after the cessation of probiotic supplementation.
CONCLUSION:
L. salivarius LS01 seems to be able to improve the quality of life of children affected by AD and, as a consequence, it may have promising clinical and research implication.
Treatment of atopic dermatitis eczema with a high concentration of Lactobacillus salivarius LS01 associated with an innovative gelling complex: a pilot study on adults.
Drago L 1, De Vecchi E, Toscano M, Vassena C, Altomare G, Pigatto P.
ABSTRACT:
GOAL:
To evaluate the efficacy of a highly concentrated Lactobacillus salivarius preparation containing a gelling complex formed by Streptococcus thermophilus ST10 and tara gum in the treatment of atopic dermatitis (AD).
BACKGROUND:
Previous studies have demonstrated an improvement in AD symptoms after administration of the probiotic strain L. salivarius LS01. S. thermophilus ST10 and tara gum create a gelling complex that adheres to intestinal mucus and improves barrier function.
STUDY:
A prospective, controlled pilot trial was carried out to evaluate how the association of S. thermophilus ST10 and tara gum could improve the activity of L. salivarius LS01 administered at high doses to adults with AD. Twenty-five patients were included into the study: 13 were treated for 1 month with the active formulation, whereas 12 represented the placebo group. Scoring Atopic Dermatitis index was determined before and at the end of probiotic administration. Fecal samples were also collected to evaluate changes in bacterial counts of Staphylococcus aureus and clostridia.
RESULTS:
A significant improvement in SCORAD index was observed in the probiotic group after 1 month of treatment, whereas no significant changes occurred in placebo patients. A slight decrease in fecal S. aureus count was observed in probiotic-treated patients.
CONCLUSION:
Data obtained in this study suggest a potential role for L. salivarius LS01 in the treatment of AD. The addition of tara gum and S. thermophilus ST10 seems to improve the overall efficacy of the probiotic strain, in particular shortening the time required for the onset of the positive effects. Further studies to investigate the activity of this preparation are advisable.
The innovative potential of Lactobacillus rhamnosus LR06, Lactobacillus pentosus LPS01, Lactobacillus plantarum LP01, and Lactobacillus delbrueckii Subsp. delbrueckii LDD01 to restore the "gastric barrier effect" in patients chronically treated with PPI: a pilot study.
Del Piano M 1, Anderloni A, Balzarini M, Ballarè M, Carmagnola S, Montino F, Orsello M, Pagliarulo M, Tari R, Soattini L, Sforza F, Mogna L, Mogna G.
ABSTRACT:
BACKGROUND:
Gastroesophageal reflux disease is a very widespread condition. In Europe, it is estimated that about 175 million people suffer from this disease and have to chronically take drugs to increase gastric pH. The proton pump inhibitors (PPIs) such as omeprazole, lansoprazole, and esomeprazole are the most widely used drug typology in this regard. However, the inhibition of normal gastric acid secretion has important side effects, the most important being bacterial overgrowth in the stomach and duodenum with a concentration of >10⁵ viable cells/mL. As a major consequence of this, many harmful or even pathogenic bacteria contained in some foods could survive the gastric transit and colonize either the stomach itself, the duodenum, or the gut, where they could establish acute and even chronic infections with unavoidable consequences for the host’s health. In other words, the "gastric barrier effect" is strongly reduced or even disrupted. To date, there are no real strategies to deal with this widespread, although still relatively little known, problem. The aim of this study was to confirm the gastric bacterial overgrowth in long-term PPI consumers and to assess the efficacy of some probiotic bacteria, belonging to both genera Lactobacillus and Bifidobacterium, in the reduction of gastric and duodenal bacterial overgrowth, therefore partially restoring the gastric barrier effect against foodborne pathogenic bacteria.
METHODS:
For this purpose, probiotics with a strong demonstrated inhibitory activity on gram-negative bacteria, such as Escherichia coli, were tested in a human intervention trial involving a total of 30 subjects treated with PPIs for either 3 to 12 consecutive months (short-term) or >12 consecutive months (long-term). An additional 10 subjects not taking PPIs were enrolled and used as a control group representing the general population. Four selected probiotics Probiotical SpA (Novara, Italy), namely Lactobacillus rhamnosus LR06 (DSM 21981), Lactobacillus pentosus LPS01 (DSM 21980), Lactobacillus plantarum LP01 (LMG P-21021), and Lactobacillus delbrueckii subsp. delbrueckii LDD01 (DSM 22106) were administered for 10 days to 10 subjects treated with PPIs for >12 months (group B). In the 60 mg formulation, N-acetylcysteine was included as well in light of its well-known mechanical effects on bacterial biofilms. Gastroscopies were performed at the beginning of the study (d0) in all the groups (A, B, C, and D) and after 10 days (d10) in group B only; that is, at the end of probiotics intake. The total viable cells and total Lactobacillus were quantified in gastric juice and duodenal brushing material from all subjects. The results were compared among all the groups and with the control subjects (group D) to confirm the bacterial overgrowth. A comparison was made also between d0 and d10 in group B to quantify the efficacy of the 4 probiotics administered for 10 days. Fecal samples were collected from all groups at d0, including subjects not treated with PPIs, and in group B only at d10. Specific bacterial classes, namely enterococci, total coliforms, E. coli, molds, and yeasts were quantified in all fecal specimens.
RESULTS:
The results collected confirmed the strong bacterial overgrowth in the stomach and duodenum of people treated with PPIs compared with subjects with a normal intragastric acidity. It is also worth noting that the bacterial cell counts in subjects who underwent a long-term treatment with a PPI were greater than the results from subjects taking these drugs for 3 to 12 months. The intake of 4 specific probiotic strains with a marked antagonistic activity towards 5 E. coli bacteria, including the enterohaemorrhagic O157:H7 strain, and an effective amount of N-acetylcysteine (NAC) was able to significantly reduce bacterial overgrowth in long-term PPI-treated subjects. Total lactobacilli represented the major percentage of bacterial counts, thus demonstrating the ability of such bacteria to colonize the stomach and the duodenum, at least temporarily, and to consequently restore the gastric barrier effect. A significant decrease in fecal enterococci, total coliforms, E. coli, molds, and yeasts in subjects treated with PPIs was recorded at the end of probiotics supplementation (d10) compared with baseline (d0) in group B. This is a further confirmation of the barrier effect also exerted at the stomach level.
CONCLUSION:
PPIs are the most widely sold and used drugs in the world. However, the chronic use of these pharmacological molecules exposes the subject to the risk of foodborne infections as most pathogens are able to survive the gastric transit in a condition of significantly decreased acidity.
Probiotics in the treatment of irritable bowel syndrome.
Saggioro A.
ABSTRACT:
Irritable Bowel Syndrome (IBS) may be diagnosed on the presence of symptoms, according to Rome II criteria, [corrected] and some studies have shown that abnormal colonic fermentation may be an important factor in the development of symptoms in some patients with IBS. Since the fermentation [corrected] of substrates by the intestinal flora may play a key role in the use of probiotics in the treatment of IBS, seventy [corrected] patients (31 [corrected] males, 39 [corrected] females), mean age 40 years (range = 26-64 years) with IBS, according to Rome II criteria, were enrolled into the study after informed consensus. Patients were randomly assigned to receive for 4 weeks [corrected] either the active preparation containing Lactobacillus plantarum LP 01 [corrected] and Bifidobacterium breve BR 03 [corrected] or Lactobacillus plantarum LP 01 and Lactobacillus acidophilus LA 02, all strains at concentrations of 5 x 10(9) CFU/g) [corrected] or placebo powder containing starch identical to the study product [corrected] To evaluate treatment efficacy two different scores were considered [corrected] Pain score in different abdominal locations after treatment decreased in probiotics groups A and B 42% and 49% versus 25% [corrected] (P < 0.05) in [corrected] placebo group after 14 days and 45% and 49% versus 29.5% [corrected] (P < 0.001) after 28 days. The severity score of characteristic IBD symptoms significantly decreased in probiotic groups A and B [corrected] versus placebo group after 14 days, 49.3% and 55.6% [corrected] versus 8% [corrected] (P < 0.001), and these data were confirmed after 28 days (56% and 55.6% versus 14.4% [corrected] P < 0.001). In conclusion, short-term therapy with Lactobacillus plantarum LP 01 and Bifidobacterium breve BR 03 or Lactobacillus plantarum LP 01 and Lactobacillus acidophilus LA 02 [corrected] may be considered a promising approach for IBS therapy [corrected]
Multi strain symbiotic preparations as a Novel Adjuvant Approach to Allergic Rhinitis
Assessment of the capability of a gelling complex made of tara gum and the exopolysaccharides produced by the microorganism Streptococcus thermophilus ST10 to prospectively restore the gut physiological barrier: a pilot study.
Del Piano M 1, Balzarini M, Carmagnola S, Pagliarulo M, Tari R, Nicola S, Deidda F, Pane M.
ABSTRACT:
BACKGROUND:
Leaky gut, or intestinal permeability, is the phenomenon of the gut wall exhibiting increased absorbency. It is pretty well recognised that an altered or damaged bowel lining or gut wall may result from unbalanced diet, parasites, infection, or medications and that this allows substances such as toxins, microbes, undigested food, or waste to leak through. As a natural consequence, this prompts the body to initiate an immune reaction leading to potentially severe health conditions. Different strategies may be used to improve, at least temporarily, the physiological intestinal barrier. The use of specific beneficial microorganisms, such as lactobacilli and bifidobacteria, has been suggested as an innovative tool to counteract an improper level of intestinal permeability. The association of bacteria with specific gelling agents, such as gums, may represent an improvement since these molecules are able to form hydrophilic gels that distribute uniformly over the inner intestinal surface. This pilot study was undertaken to evaluate intestinal permeability in subjects treated with a gelling complex, an association of tara gum and the microorganism Streptococcus thermophilus ST10 (DSM 25246), which has a well-demonstrated in vitro ability to synthesise and secrete exopolysaccharides (EPSs).
METHODS:
Twenty-five healthy subjects were enrolled in this human intervention, double-blind, placebo-controlled, pilot trial (age between 21 and 57 y, mean 37.7±11.2). Subjects were then randomised into 2 groups: group A (13 subjects) was given an active formulation containing 250 mg of tara gum and 1 billion viable cells of S. thermophilus ST10, whereas group B (12 subjects) was given a placebo formulation. All the subjects participating in the study were directed to take 1 dose per day for 30 consecutive days. The presence and concentration of exopolysaccharides (EPSs) in the faeces was determined at time 0 (d0), after 30 days of treatment (d30), and at the end of the 2-week follow-up period (d45). The monosaccharide composition of EPSs was used to quantify the possible contribution of tara gum to the amount of polysaccharides detected in the faecal material. Intestinal permeability was evaluated at the same time by means of the lactitol/mannitol ratio (small intestine permeability) and sucralose concentration (colonic permeability) in urine specimens sampled after specified times. A statistical comparison was made between the concentration of EPSs, the lactulose/mannitol ratio, and the amount of excreted sucralose in the 2 groups at d0, d30, and d45.
RESULTS:
In the active group, supplementation with S. thermophilus ST10 and tara gum was able to significantly increase the faecal EPSs concentration compared with placebo (from 0.169 mg/g to 0.633 mg/g after 30 d, P<0.001). An interesting decrease in intestinal permeability, both of the small bowel and in the colon, was also recorded. The L/M ratio diminished from 0.021 in the active group to 0.014 and 0.015 after 30 and 45 days, respectively (P=0.045 and P=0.033 compared with placebo). The sucralose concentration decreased from 35.8 mg to 27.9 mg and 29.1 mg (P=0.038 and P=0.026 compared with placebo) at the end of the supplementation period and after the follow-up, respectively. No significant differences were recorded in the placebo after 30 days or at the end of the follow-up.
CONCLUSION:
The association of the EPSs produced by S. thermophilus ST10 and tara gum seems capable of significantly improving the intestinal functional barrier in healthy subjects. A wider study in subjects presenting impaired gut permeability would be useful in the future to confirm the positive results from this pilot trial. In any case, our findings are consistent with the parallel increase in exopolysaccharide concentration in the faecal material, thus suggesting the effective ability of the strain used to secrete EPSs in the gut lumen. An innovative approach of this type may be useful in helping to restore the physiological barrier by means of a merely natural and mechanical action.
Comparison of the kinetics of intestinal colonization by associating 5 probiotic bacteria assumed either in a microencapsulated or in a traditional, uncoated form.
Piano MD 1, Carmagnola S, Ballarè M, Balzarini M, Montino F, Pagliarulo M, Anderloni A, Orsello M, Tari R, Sforza F, Mogna L, Mogna G.
ABSTRACT:
BACKGROUND:
Beneficial findings concerning probiotics are increasing day by day. However, one of the most important parameters able to significantly affect the probiotic value of a microorganism is its survival during the transit through the stomach and the duodenum. Some techniques may be applied that aim to improve this parameter, but microencapsulation of bacterial cells remains one of the most important. A recent study assessed the kinetics of intestinal colonization by a mixture of 2 probiotic strains, given either in a microencapsulated or in a traditional, uncoated form.
METHODS:
A comparison between the intestinal colonization by associating 5 microencapsulated bacteria and the same uncoated strains was performed by a double-blind, randomized, cross-over study. The study (December 2007 to January 2009) involved 53 healthy volunteers. In particular, subjects were divided into 2 groups: group A (27 subjects) was given a mix of probiotic strains Probiotical S.p.A. (Novara, Italy), Lactobacillus acidophilus LA02 (DSM 21717), Lactobacillus rhamnosus LR04 (DSM 16605), L. rhamnosus GG, or LGG (ATCC 53103), L. rhamnosus LR06 (DSM 21981), and Bifidobacterium lactis BS01 (LMG P-21384) in an uncoated form, whereas group B (26 subjects) received the same strains microencapsulated with a gastroprotected material. The uncoated strains were administered at 5×10⁹ cfu/strain/d (a total of 25×10⁹ cfu/d) for 21 days, whereas the microencapsulated bacteria were given at 1×10⁹ cfu/strain/d (a total of 5×10⁹ cfu/d) for 21 days. At the end of the first period of supplementation with probiotics, a 3-week wash-out phase was included in the study setting. At the end of the wash-out period, the groups crossed over their treatment regimen; that is, group A was administered the microencapsulated bacteria and group B the uncoated bacteria. The administered quantities of each strain were the same as the first treatment. A quantitative evaluation of intestinal colonization by probiotics, either microencapsulated or uncoated, was undertaken by examining fecal samples at the beginning of the study (time 0), after 10 days and after 21 days of each treatment period. In particular, fecal total Lactobacilli, heterofermentative Lactobacilli, and total Bifidobacteria were quantified at each checkpoint. A genomic analysis of an appropriate number of colonies was performed to quantify individual L. rhamnosus strains among heterofermentative Lactobacilli.
RESULTS:
A statistically significant increase in the fecal amounts of total Lactobacilli, heterofermentative Lactobacilli, and total Bifidobacteria was registered in both groups at the end of each supplementation period compared with d₀ or d₄₂ (group A: P=0.0002, P=0.0001, and P<0.0001 at d₂₁, P=0.0060, P=0.0069, and P<0.0001 at d₆₃ for total Lactobacilli, heterofermentative Lactobacilli, and Bifidobacteria, respectively; group B: P=0.0002, P=0.0006, and P<0.0001 at d₂₁, P=0.0015, P=0.0016, and P<0.0001 at d₆₃ for total Lactobacilli, heterofermentative Lactobacilli, and Bifidobacteria, respectively), confirming the ability of each strain in the administered composition to colonize the human gut, whether supplemented in a gastroprotected or in a traditional freeze-dried form. On the contrary, subjects receiving microencapsulated bacteria reported a kinetics of intestinal colonization that was entirely comparable with those who were given uncoated strains at a 5 times higher amount.
CONCLUSION:
The microencapsulation technique used in this study is a valid approach aimed to significantly improve the survival of strains during gastroduodenal transit, thus enhancing their probiotic value and allowing the use of a 5 times lower amount.
Evaluation of the intestinal colonization by microencapsulated probiotic bacteria in comparison with the same uncoated strains.
Del Piano M 1, Carmagnola S, Andorno S, Pagliarulo M, Tari R, Mogna L, Strozzi GP, Sforza F, Capurso L.
ABSTRACT:
BACKGROUND:
Beneficial findings concerning probiotics are increasing day by day. However, one of the most important parameter which affects the probiotic activity of a microorganism is its survival during the gastroduodenal transit. Some microencapsulation techniques could be applied to bacterial cells to improve this parameter.
METHODS:
A comparison between the intestinal colonization by microencapsulated bacteria and the same not microencapsulated strains has been conducted in a double blind, randomized, cross-over study. The study (April to July 2005) involved 44 healthy volunteers. In particular, participants were divided into 2 groups: group A (21 participants) received a mix of probiotic strains Lactobacillus plantarum LP01 (LMG P-21021) and Bifidobacterium breve BR03 (DSM 16604) in an uncoated form, group B (23 participants) was given the same strains microencapsulated with a gastroresistant material. The not microencapsulated strains were administered at 5 x 10(9) colony forming units/strain/d for 21 days, whereas the microencapsulated bacteria were given at 1 x 10(9) colony forming units/strain/d for 21 days. At the end of the first period of treatment with probiotics a 3 weeks washout phase has been included in the study protocol. At the end of the washout period the groups were crossed: in detail, group A had the microencapsulated and group B the uncoated bacteria. The administered amounts of each strain were the same as the first treatment. The quantitative evaluation of intestinal colonization by strains microencapsulated or not microencapsulated was made by fecal samples examination at the beginning of the clinical trial, after 10 and 21 days of each treatment period. In particular, fecal heterofermentative Lactobacilli and Bifidobacteria have been counted.
RESULTS:
A statistically significant increase in the fecal amounts of Lactobacilli and Bifidobacteria was recorded in both groups at the end of each treatment compared with d0 or d42 (P<0.0001 and P<0.0001 at d21, P<0.0001 and P<0.0001 at d63 for Lactobacilli and Bifidobacteria, respectively), confirming the ability of the 2 strains to colonize the human gut, either in a gastroprotected form or not. Participants treated with the microencapsulated bacteria reported a kinetics of intestinal colonization quite similar to participants who received not coated strains.
CONCLUSION:
Probiotics are able to exert many different beneficial effects on the human host. These effects are mediated by the number of viable cells which reach the gut. The microencapsulation technique used in this study is a valid strategy to significantly improve gastroresistance of strains, thus enhancing their probiotic activity and allowing the use of a 5 times lower amount.
Screening of different probiotic strains for their in vitro ability to metabolise oxalates: any prospective use in humans?
Mogna L 1, Pane M, Nicola S, Raiteri E.
ABSTRACT:
BACKGROUND:
Oxalate is the salt-forming ion of oxalic acid and can generate oxalate salts combining with various cations, such as sodium, potassium, magnesium, and calcium. Approximately 75% of all kidney stones are composed primarily of calcium oxalate (CaOx) and hyperoxaluria, a condition involving high urinary oxalate concentration, is considered a primary risk factor for kidney stone formation, known as nephrolithiasis. Current therapeutic strategies often fail in their compliance or effectiveness, and CaOx stone recurrence is still common. After an initial stone, there is a 50% chance of forming a second stone within 7 years if the condition is left untreated. The potential therapeutic application of some probiotics, mainly lactobacilli and bifidobacteria, in reducing hyperoxaluria in vivo through intestinal oxalate degrading activity is compelling and initial reports are promising. This study was undertaken to screen different Lactobacillus and Bifidobacterium strains for their capacity to degrade oxalate in vitro using reverse-phase high-performance liquid chromatography (HPLC).
METHODS:
The oxalate-degrading activity of 13 lactobacilli and 5 bifidobacteria was tested using a novel HPLC method after growth in a broth culture added with 10 mM ammonium oxalate. Experiments were repeated 3 times. Oxalobacter formigenes (DSM 4420) was used as positive reference to validate HPLC oxalate-degrading capability assays.
RESULTS:
Lactobacillus strains were more efficient than bifidobacteria in degrading oxalates. L. paracasei LPC09 (DSM 24243) gave the best result, as 68.5% of ammonium oxalate was converted at the end of incubation, whereas the following best converters belong to the L. gasseri and L. acidophilus species. The relatively low conversion rate observed for most bifidobacteria can probably be attributed to intrinsic oxalate toxicity toward this genus.
CONCLUSION:
Humans lack the enzymes needed to directly metabolise oxalate, and this potentially toxic compound is, therefore, managed using alternative pathways. As oxalate-degrading bacteria are present in the endogenous microbiota of the human intestine, although with significant individual differences, it is possible to hypothesise that the administration of selected oxalate-degrading probiotics could be an alternative and innovative approach to reducing the intestinal absorption of oxalate and the resulting urinary excretion.
Probiotics as a Novel Adjuvant Approach to Atopic Dermatitis
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