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Probiotic agents to protect the urogenital tract against infection.

The urogenital microflora of a healthy woman comprises approximately 50 species of organisms, which differ in composition according to reproductive stages and exposure to several factors, including antibiotics and spermicides. Infections are very common with > 300 million cases of urinary tract infections, bacterial vaginosis, and yeast vaginitis worldwide per annum. At the time of infection in the bladder and vagina, the urogenital flora is often dominated by the infecting pathogens, in contrast with healthy phases when indigenous organisms dominate. Premenopausal women have a flora of mostly lactobacilli, and certain properties of these strains, including adhesive ability and production of acids, bacteriocins, hydrogen peroxide, and biosurfactants, appear important in conferring protection to the host. Efforts to artificially restore an unbalanced flora with the use of probiotics have met with mixed results but research aimed at selecting scientifically based strains could well provide a reliable alternative treatment and preventive regimen to antibiotics in the future.

Anti-Bacterial Agents↗

Enhancement of natural killer cytotoxicity delayed murine carcinogenesis by a probiotic microorganism.

Regulation of innate immunity may be an effective means of cancer control. Delaying cancer onset is regarded as an important mode of action in cancer prevention. We have been investigating the chemopreventive mechanisms of Lactobacillus casei Shirota (LcS), a probiotic strain. In this study, we evaluated the effect of LcS on tumor onset and the involvement of natural killer (NK) cells using a 3-methylcholanthrene-induced carcinogenesis model. C3H/HeN mice were divided into three groups, according to treatment: vehicle-treated, treated with vehicle only; control, 3-methylcholanthrene treated; LcS, 3-methylcholanthrene and LcS treated. 3-Methylcholanthrene was injected intradermally at 7 weeks of age. LcS was mixed into the diet (0.05%, w/w), which the mice were fed from the day of 3-methylcholanthrene injection onward. Tumor incidence was observed weekly. Profiles of splenic NK cells, in vitro cytotoxicity and the proportion, in the early stage of carcinogenesis were analyzed at 5 weeks after the injection. The tumor suppressive effect of LcS was also evaluated in a beige mouse model that is genetically deficient in NK cells. LcS delayed tumor onset and reduced tumor incidence in the results with C3H/HeN mice (P< 0.05). More specifically, tumor incidence in the control group was 33% at 6 weeks after the injection and 83% at 11 weeks as opposed to 0 and 42%, respectively, in the LcS group. NK cell cytotoxicity was significantly higher than in the control group, and the number of NK cells also increased in the LcS group of C3H/HeN mice. However, LcS failed to suppress tumorigenesis in the beige mouse. These findings suggest that enhancement of the cytotoxicity of NK cells by LcS delays tumor onset.

Animals↗

Gas chromatographic method for analysis of conjugated linoleic acids isomers (c9t11, t10c12, and t9t11) in broth media as application in probiotic studies.

A gas chromatography (GC) procedure is assayed for analysis of conjugated linoleic acid (CLA) isomers cis-9, trans-11-octadecadienoic (c9t11); trans-10, 12 cis-octadecadienoic (t10c12); and trans-9, trans-11-octadecadienoic (t9t11) in culture broth by GC using NaOH-BF3 in methanol for methylating and a long capillary (100 m) high-polarity column. Repeatability of the method is assessed; the coefficient of variation for CLA isomers ranges from 4.62 for c9t11 to 8.19 for t9t11. Recovery ranges between 88.01 and 89.76, with a mean value of 89.06 for all CLA isomers studied. This method may be considered advantageous for analysis of CLA isomers in probiotics cultures samples.

Chromatography, Gas↗

Aspects of in vitro and in vivo research approaches directed toward identifying probiotics and prebiotics for human use.

The microbiota of the human gastrointestinal tract plays a key role in nutrition and health. Through the process of fermentation, gut bacteria metabolize various substrates (principally dietary components) to end products such as short-chain fatty acids and gases. This anaerobic metabolism is thought to contribute positively toward host daily energy requirements. However, under certain circumstances, the fermentative process may produce undesirable metabolites. This may cause the onset of gut disorders that can be manifest through both acute and chronic conditions. Moreover, the gut flora may become contaminated by transient pathogens that serve further to upset the normal community structure. There has been a recent increase in the use of dietary components that help to maintain, or even improve, the gut microflora "balance." Probiotics are live microbial feed supplements added to appropriate food vehicles (usually fermented milks), whereas prebiotics are dietary carbohydrates that have a selective metabolism in the colon and serve to increase numbers of bacteria seen as desirable. Because of their purported health-promoting properties, lactic acid-producing bacteria, including bifidobacteria, are the usual target organisms. The market value and biological potential of both approaches are enormous. This article will summarize how efficacious types can be identified.

Fermentation↗

Probiotic bacteria: today and tomorrow.

This paper provides an overview of the key issues raised during this symposium. Probiotic cultures have been associated historically with cultured milks and dairy products, from which there is substantial evidence for positive effects on human health and general well-being.

Animals↗

Prebiotic inulin enriched with oligofructose in combination with the probiotics Lactobacillus rhamnosus and Bifidobacterium lactis modulates intestinal immune functions in rats.

Probiotics (PRO) modulate systemic immunity in animals and humans. In contrast, the effects of prebiotics (PRE) on systemic and intestinal immunity have not been investigated. Whether the combined application of PRO and PRE [synbiotics (SYN)] has synergistic or additive effects is presently unknown. Therefore, PRO (Lactobacillus rhamnosus GG and Bifidobacterium lactis Bb12), PRE (inulin enriched with oligofructose), and SYN (combination of PRO and PRE) were fed to F344 rats for 4 wk as supplements to a high fat diet. Functions of immune cells isolated from peripheral blood mononuclear cells (PBMC), spleen, mesenterial lymph nodes and Peyer's patches (PP) were investigated. The SYN supplement increased secretory immunoglobulin A (sIgA) production in the ileum compared with controls fed the high fat diet alone (P<0.05), and decreased the oxidative burst activity of blood neutrophils (P<0.05) compared with rats fed PRO. The PRE supplement enhanced the production of interleukin-10 (P<0.05) in PP as well as the production of sIgA in the cecum (P<0.05), compared with controls. The PRO supplement modestly affected immune functions, whereas systemic immunomodulatory effects were observed in rats fed SYN. The PRE supplement primarily acted at the level of the gut-associated lymphoid tissue. The combined application of PRO and PRE has different effects from those of the individual supplements, but does not simply result in additive or synergistic effects.

Animals↗

Supplementation of infant formula with the probiotic lactobacillus reuteri and zinc: impact on enteric infection and nutrition in infant rhesus monkeys.

UNLABELLED: Gut colonization by may have beneficial effects on infant health or capacity to resist infectious disease. Zinc supplementation has also been proposed to increase infants' resistance to disease; however, many studies have yielded conflicting results. OBJECTIVES: To study effects of probiotic supplementation of infant formula (with or without supplemental zinc) on nutritional status, gut colonization and the ability to resist gastrointestinal infection in an infant rhesus monkey model. METHODS: Infant monkeys were fed control infant formula (5 mg Zn/L), control formula with or control formula with and supplemental zinc (15 mg Zn/L) from birth to 4 months. Growth, nutritional status, mineral absorption, intestinal colonization and frequency and severity of enteropathogenic -induced gastroenteritis were monitored. RESULTS: Gastrointestinal colonization was achieved and was associated with increased ileal villous surface area and improved hematocrit, with no adverse effects on growth or nutritional indices. Fortification to 15 mg Zn/L reduced plasma copper, erythrocyte Cu/Zn-superoxide dismutase, hemoglobin, and iron absorption. Infants fed -supplemented formula had reduced diarrhea severity throughout the study period and recovered more rapidly from acute diarrhea than the other groups. CONCLUSION: -supplementation of infant formula is safe, improves iron status and decreases diarrhea severity in infant rhesus monkeys and thus may help protect formula-fed human infants from infection and nutritional deficiencies.

Animals↗

Probiotic bacteria in the management of atopic disease: underscoring the importance of viability.

OBJECTIVES: The aim of this study was to assess the efficacy of oral supplementation of viable and heat-inactivated probiotic bacteria in the management of atopic disease and to observe their effects on the composition of the gut microbiota. METHODS: The study population included 35 infants with atopic eczema and allergy to cow's milk. At a mean age of 5.5 months, they were assigned in a randomized double-blind manner to receive either extensively hydrolyzed whey formula (placebo group) or the same formula supplemented with viable (viable LGG group) or heat-inactivated Lactobacillus GG (heat-inactivated LGG group), respectively. The changes in symptoms were assessed by the SCORAD method and the presence of some predominant bacterial genera in the feces detected with 16S rRNA-specific probes. RESULTS: The treatment with heat-inactivated LGG was associated with adverse gastrointestinal symptoms and diarrhea. Consequently, the recruitment of patients was stopped after the pilot phase. Within the study population, atopic eczema and subjective symptoms were significantly alleviated in all the groups; the SCORAD scores (interquartile range) decreased from 13 (range, 4-29) to 8 (range, 0-29) units in the placebo group, from 19 (range, 4-47) to 5 (range, 0-18) units in the viable LGG group, and from 15 (range, 0-29) to 7 (range, 0-26) units in the heat-inactivated LGG group. The decrease in the SCORAD scores within the viable LGG group tended to be greater than within the placebo group. The treatments did not appear to affect the bacterial numbers within the genera enumerated. CONCLUSIONS: Supplementation of infant formulas with viable but not heat-inactivated LGG is a potential approach for the management of atopic eczema and cow's milk allergy.

Administration, Oral↗

Gut microbial ecology in critical illness: is there a role for prebiotics, probiotics, and synbiotics?

Approximately 70% of the immune system is localized in the gastrointestinal tract: its glands, mucosa, and mucosa-associated lymphoid system. The system influences health conditions because it produces large amounts of important gastrointestinal secretions as rich as breast milk in health-supporting and disease-preventing factors, and because of its rich gastrointestinal flora. The intestine normally contains 10 times more microbes than there are eukaryotic cells in the entire body. The optimal function of these microbes depends on the supply of food destined for the colonic bacteria (fermentable fibers, complex proteins, gastrointestinal secretions). The consideration of these functions influences outcome. Unfortunately, the conditions (supply of drugs-especially antibiotics, and reduced supply of food-especially fruits and vegetables) in the modern ICU are extremely poor both for optimal gastrointestinal secretion and for flora and need more attention. To improve treatment, a supply of new and effective flora (probiotics) and food for the flora (prebiotics) is needed, from which numerous health-supporting products (synbiotics) will be produced and absorbed at the level of the mucosa, mainly in the lower gastrointestinal tract.

Critical Care↗

Probiotics: a potential target for the prevention and treatment of steatohepatitis.

The accumulation of fat in hepatocytes with a necroinflammatory component-steatohepatitis-that may or may not have associated fibrosis is becoming a frequent lesion. Although steatohepatitis is currently recognized to be a leading cause of cryptogenic cirrhosis, the pathogenesis has not been fully elucidated. Among the various factors implicated, intestinal bacterial overgrowth may play a role. Indeed, various rat models of intestinal bacterial overgrowth have been associated with liver lesions similar to NASH, and bacterial overgrowth has been observed significantly more often in patients with NASH compared with control subjects. The authors discuss the relationship among intestinal bacterial overgrowth, steatohepatitis development, and probiotic treatment.

Animals↗

Clinical experience with probiotics in the elderly on total enteral nutrition.

BACKGROUND: Recent data support that after 2 years of age, intestinal microflora remains relatively constant over time, except in elderly people, who harbor fewer bifidobacteria and a higher population of fungi and enterobacteria than young adults. Diet supplementation with probiotics may improve the nutritional status and reduce the impaired immunity associated with aging. The goal of this study was to establish the effect on bifidobacteria fecal counts, and some clinical parameters, of bifidobacteria supplementation to elderly patients in total parenteral nutrition. METHODS: Thirteen patients (6 men and 7 women; mean age, 69 years; range, 65-76 years) affected by permanent vegetative status (PVS) and fed by total enteral nutrition (TEN) were studied. Bifidobacteria and clostridia were investigated by microbiologic and molecular biology methods in stool specimens collected twice at basal time (T-2 and T0) and after 12 and 15 days (T12 and T15, respectively). Seven patients with basal bifidobacteria counts less than 10 were supplemented with Bifidobacterium longum W 11 for 12 days. The remaining 6 patients were used as control subjects. For 1 month diarrhea and fever episodes, use of antibiotics, and nutritional status (BMI) were assessed. RESULTS: In the 7 patients with bifidobacteria counts less than 10, the administration of B. longum W 11 resulted in a 1 log increase in 6 of 7 patients at T12. No statistically significant difference in episodes of fever or diarrhea, use of antibiotics, or BMI was observed between the treatment and control groups. CONCLUSION: The administration of B. longum W11 in PVS patients fed by TEN is effective in increasing the population of bifidobacteria. Larger studies with longer follow-up could demonstrate the influence of these microbiologic changes in a clinical setting.

Aged↗

Utilization of the intestinal tract as a delivery system for urogenital probiotics.

Orally consumed viable bacteria with proposed beneficial health effects, the so-called probiotics, are increasingly used to treat disorders of the gastrointestinal tract. It has been recently suggested that specifically selected lactobacilli could have a therapeutic role in female urogenital tract infections. It has been also suggested that some of these bacteria could be not intravaginally instilled, but also orally delivered. The authors investigate the ability of lactobacilli isolated from the vagina to survive gastrointestinal tract transit, and establish a link between the rate of intestinal survival and vaginal colonization.

Administration, Oral↗

Probiotics and functional abdominal bloating.

Functional abdominal bloating is a condition dominated by a feeling of abdominal fullness or bloating and without sufficient criteria for another functional gastrointestinal disorder. The currently used therapeutic approaches aim to reduce the volume of intestinal gas, thus increasing intestinal gas elimination or reducing its production. Some promising results have been obtained by the use of prokinetics, such as tegaserod and Prostigmine, and by the use of nonabsorbable antibiotics, such as rifaximin. Another therapeutic approach is represented by the administration of probiotics to modify the composition of colonic flora and thus the production of intestinal gas. The authors recently studied the effect of LGG, which proved to be more effective than placebo in reducing the severity of symptoms.

Abdomen↗

Probiotics for the developing world.

Every minute of every day more and more children die of diarrheal diseases and women, and girls become infected by HIV. An estimated 7,000 women become infected each day. While many valiant efforts are being made to address these issues, until now they have proved to be markedly ineffective. The notion that lactic acid bacteria, formulated into food or dietary supplements, could have a role to play in slowing the morbidity and mortality associated with HIV/AIDS and gastroenteritis, is built upon sound clinical findings and scientific investigations, yet no international efforts have been placed in this approach, to date. We hereby summarize the reasons why such efforts should be made, provide an example of one model being set up in sub-Saharan Africa, and challenge the international community to consider the potential benefits of probiotics, especially for communities not reached by governmental and nongovernmental agencies.

Developing Countries↗

Growth of probiotic lactobacilli in the presence of oleic acid enhances subsequent survival in gastric juice.

The effect of inclusion of various C18 fatty acids with 0-2 double bonds in either cis or trans configuration on Lactobacillus rhamnosus GG survival was analysed in simulated gastric juice at pH 2.5. The incorporation of Tween 80 (1 g l-1) in the growth media enhanced subsequent survival of stationary-phase cultures up to 1000-fold following 90 min acid exposure compared with controls grown without Tween 80. There was a significant (P<0.05) increase in bacterial content of oleic acid [C18:1 (9c), up to 55-fold] after growth of bacteria in MRS supplemented with Tween 80. The inclusion of various C18 fatty acids in the growth media revealed that only oleic and vaccenic acids [C18:1 (11t)] had protective effects on the survival of Lb. rhamnosus GG when exposed to the acidic environment. Comparative analysis with other lactobacilli indicated that all strains exhibited increased survival when grown in the presence of Tween 80. Further work with a neomycin-resistant mutant with 48% of the F0F1-ATPase activity of the parent indicated that the Tween 80 effect was independent of the complex. The mechanisms behind the effect of fatty acid protection were investigated and proton permeability assays showed that cultures grown in the presence of Tween 80 had higher extracellular pH than controls. Furthermore, there was a significant reduction of oleic acid and a significant increase in stearic acid (C18:0) (P<0.05) content of bacterial cells following exposure of Tween 80-supplemented cultures to simulated gastric juice. Overall, the data suggest that probiotic lactobacilli can use an exogenous oleic acid source to increase their acid survival and the underlying mechanism most likely involves the ability of increased membrane oleic acid to be reduced by H+ to stearic acid.

Culture Media↗

Use of a probiotic to control lactococcosis and streptococcosis in rainbow trout, Oncorhynchus mykiss (Walbaum).

From a comparison of 125 bacterial isolates recovered from the digestive tract of rainbow trout, Oncorhynchus mykiss, and carp, Cyprinus sp., a culture was obtained which was effective at preventing clinical disease caused by Lactococcus garvieae and Streptococcus iniae when used as a feed additive. The culture, Aeromonas sobria GC2, was incorporated into the feed and fed to rainbow trout (average weight = 20 g) for 14 days at a dose equivalent to 5 x 10(7) cells g(-1) of feed. Whereas the untreated controls experienced losses of 75-100% when challenged intraperitoneally with L. garvieae and S. iniae, the probiotic-treated groups remained healthy with total mortalities of only 0-6%. Formalized and sonicated preparations of GC2 and cell-free supernatant fared less well. The mode of action reflected stimulation of innate immunity, namely an increased number of leucocytes and enhanced phagocytic and respiratory burst activity.

Aeromonas↗

A randomized controlled trial of a probiotic combination VSL# 3 and placebo in irritable bowel syndrome with bloating.

AIM: To evaluate the effects of a combination probiotic on symptoms and colonic transit in patients with irritable bowel syndrome (IBS) and significant bloating. METHODS: Forty-eight patients with Rome II IBS were randomized in a parallel group, double-blind design to placebo or VSL# 3 twice daily (31 patients received 4 weeks and 17 patients 8 weeks of treatment). Pre- and post-treatment colonic transit measurements were performed using scintigraphy with (111)In charcoal. Symptoms were summarized as an average daily score for the entire period of treatment and separately for the first 4 weeks of treatment. Weekly satisfactory relief of abdominal bloating was assessed. RESULTS: Treatment with VSL# 3 was associated with reduced flatulence over the entire treatment period (placebo 39.5 +/- 2.6 vs VSL# 3 29.7 +/- 2.6, P = 0.011); similarly, during the first 4 weeks of treatment, flatulence scores were reduced (placebo 40.1 +/- 2.5 vs VSL# 3 30.8 +/- 2.5, P = 0.014). Proportions of responders for satisfactory relief of bloating, stool-related symptoms, abdominal pain and bloating scores were not different. Colonic transit was retarded with VSL# 3 relative to placebo (colon geometric center 2.27 +/- 0.20 vs 2.83 +/- 0.19, P = 0.05 respectively). CONCLUSION: VSL# 3 reduces flatulence scores and retards colonic transit without altering bowel function in patients with IBS and bloating.

Adult↗

Field evaluation of the efficacy of a probiotic containing Bacillus licheniformis and Bacillus subtilis spores, on the health status and performance of sows and their litters.

The aim of this study was to assess the efficacy of BioPlus 2B, a probiotic containing Bacillus licheniformis and Bacillus subtilis spores, on the health status and productivity of sows and their litters. A total of 109 gilts and sows were allocated into two experimental groups, as follows: untreated controls (UC) and BioPlus 2B (same feeding as the UC group plus BioPlus 2B) at a dose of 400 g/ton of feed (equal to 1.28 x 10(6) viable spores/g of feed). Treatment started from the day of allocation (14 days prior to the expected farrowing) up to the weaning day. Homogeneity of the groups was satisfied with regard to the parity. From the results it was evident that BioPlus 2B supplementation of the feed improved gilt/sow performance as shown by: (i) the increase of sow feed consumption during the first 14 days postpartum and (ii) the decrease of sow weight loss during the suckling period. Certain blood and milk parameters were significantly improved, as shown by higher serum cholesterol and total lipids concentrations and higher milk fat and protein content at mid-suckling period. As a consequence, a positive effect was also noticed as regard litter health and performance characteristics in terms of: (i) decrease in piglet diarrhoea score, (ii) decrease in pre-weaning mortality thus leading to increase in the number of weaned piglets per litter and (iii) increase in piglet body weight at weaning. Moreover, BioPlus 2B tended to improve the health status and fertility of sows demonstrating: (i) tendency to a lower proportion of sows with Mastitis-Metritus-Agalactia (MMA) problems and (ii) lower proportion of sows returning to oestrus.

Animal Feed↗