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Augmentation of antimicrobial metronidazole therapy of bacterial vaginosis with oral probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14: randomized, double-blind, placebo controlled trial.

This study enrolled 125 premenopausal women diagnosed with bacterial vaginosis (BV) by presence of vaginal irritation, discharge and 'fishy' odor, and Nugent criteria and detection of sialidase enzyme. The subjects were treated with oral metronidazole (500 mg) twice daily from days 1 to 7, and randomized to receive oral Lactobacillus rhamnosus GR-1 (1 x 10(9)) and Lactobacillus reuteri RC-14 (1 x 10(9)) or placebo twice daily from days 1 to 30. Primary outcome was cure of BV as determined by normal Nugent score, negative sialidase test and no symptoms or signs of BV at day 30. A total of 106 subjects returned for 30-day follow-up, of which 88% were cured in the antibiotic/probiotic group compared to 40% in the antibiotic/placebo group (p<0.001). Of the remaining subjects, 30% subjects in the placebo group and none in the probiotic group had BV, while 30% in the placebo and 12% in the probiotic group fell into the intermediate category based upon Nugent score, sialidase result and clinical findings. High counts of Lactobacillus sp. (>10(5) CFU/ml) were recovered from the vagina of 96% probiotic-treated subjects compared to 53% controls at day 30. In summary, this study showed efficacious use of lactobacilli and antibiotic in the eradication of BV in black African women.

Adolescent↗

Application of a microplate scale fluorochrome staining assay for the assessment of viability of probiotic preparations.

Cell viability in probiotic preparations is traditionally assessed by the plate count technique. Additionally, fluorescent staining combined with epifluorescence microscopy or flow cytometry has been developed for the viability assessment, but the currently available assays are either laborious or require highly sophisticated equipment. The aim of this study was to investigate the applicability of a microplate scale fluorochrome assay for predicting the cell state of freeze-dried Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis preparations. In addition to viability assessment with LIVE/DEAD BacLight Bacterial Viability Kit, DiBAC(4)3 stain was used for the kinetic measurement of changes in bifidobacterial cell membrane functions during exposure to low pH. The microplate scale fluorochrome assay results on the viability and cell numbers of probiotic preparations correlated well with the results obtained with the culture-based technique and (with few exceptions) with epifluorescence microscopy. The assay was applicable also for the viability assessment of stressed (acid-treated) cells provided that the cell density in treatments was adjusted to the optimal measurement level of the fluorometer. The microplate scale fluorochrome assay offers a rapid and robust tool for the viability assessment of probiotic preparations, and enables also kinetic measurements.

Bifidobacterium↗

The potential mechanisms involved in the anti-carcinogenic action of probiotics.

Probiotic bacteria are live microbial food ingredients that provide a health benefit to the consumer. In the past it was suggested that they served to benefit the host primarily through the prevention of intestinal infections. More recent studies have implicated probiotic bacteria in a number of other beneficial effects within the host including: *The suppression of allergies. *Control of blood cholesterol levels. *Modulation of immune function. *And the prevention of cancers of the colon. The reputed anti-carcinogenic effect of probiotics arises from in vivo studies in both animals and to a limited extent in man; this evidence is supported by in vitro studies with carcinoma cell lines and anti-mutagenicity assays. However, the mechanisms involved in any effect have thus far been difficult to elucidate; studies offer evidence for a variety of mechanisms; we have reviewed these and come to the opinion that, the anti-carcinogenic effect may not be attributable to a single mechanism but rather to a combination of events not yet fully elucidated or understood.

Animals↗

Effects of probiotics on radiation-induced intestinal injury in rats.

OBJECTIVE: Radiotherapy is an important aspect of multimodal cancer therapy, but radiation-induced acute intestinal injury is a common and serious problem. Disruption of morphologic mucosal integrity and normal bacterial microflora after abdominal radiation leads to malabsorption and bacterial translocation. METHODS: Lactobacillus bulgaricus strain isolated from yogurt was given as a probiotic to rats subjected to radiotherapy. On postradiation day 8 rats were killed. Mesenteric lymph nodes, liver, and spleen were excised for microbiologic examinations. Segments of jejunum, ileum, and colon were evaluated for the presence of inflammation, vascularity, and mucus cells. RESULTS: The results of this study suggest that probiotics may have a protective effect on intestinal mucosa. CONCLUSION: Probiotics added as substrates can be given by an oral or enteral route to patients who undergo radiotherapy to prevent radiation-induced enteritis and related malnutrition.

Animals↗

Sequence analysis of the plasmid genome of the probiotic strain Lactobacillus paracasei NFBC338 which includes the plasmids pCD01 and pCD02.

Lactobacillus paracasei NFBC338 is a probiotic strain that was isolated from the human gastrointestinal tract (GIT) and contains a plasmid genome of 80kb. Using a shotgun sequencing approach, two of the plasmids, pCD01 (19,882bp) and pCD02 (8554bp) have been completely sequenced, and four contiguous sequences (Contigs) have been assembled. Bioinformatic analysis of pCD01 revealed that it contains 23 putative open reading frames (ORFs) and that it contains regions characterised by potential replication functions and multidrug resistance (MDR). In contrast, the content of pCD02 is mainly cryptic, although, it does contain two insertion sequence (IS) elements. Indeed, up to 17% of the entire plasmid genome encodes putative transposable elements. In addition, there are a number of interesting ORFs distributed over the four Contigs that show significant homology to genes such as those involved in adherence and biotin metabolism, which may prove beneficial to Lb. paracasei NFBC338 under certain environmental conditions. This study provides a novel insight into the rich plasmid complement of this probiotic Lactobacillus strain, which may potentially be exploited as the basis for development of improved genetic tools for probiotic lactobacilli.

Adhesins, Bacterial↗

Probiotic bacteria reduced duration and severity but not the incidence of common cold episodes in a double blind, randomized, controlled trial.

To investigate the effect of long-term consumption of probiotic bacteria on viral respiratory tract infections (common cold, influenza), a randomized, double blind, controlled intervention study was performed during two winter/spring periods (3 and 5 month). Four hundred and seventy-nine healthy adults were supplemented daily with vitamins plus minerals with or without probiotic lactobacilli and bifidobacteria. The intake of the probiotic had no effect on the incidence of common cold infections (verum=158, control=153 episodes, influenza was not observed), but significantly shortened duration of episodes by almost 2 days (7.0+/-0.5 versus 8.9+/-1.0 days, p=0.045), reduced the severity of symptoms and led to larger increases in cytotoxic T plus T suppressor cell counts and in T helper cell counts.

Adolescent↗

Immune responses in rainbow trout Oncorhynchus mykiss induced by a potential probiotic bacteria Lactobacillus rhamnosus JCM 1136.

This study was undertaken to examine the effect of supplementing a suggested probiotic bacteria Lactobacillus rhamnosus JCM 1136 in feed on immune response and gut flora composition of rainbow trout Oncorhynchus mykiss. The probiotic bacteria were incorporated into a commercial feed to constitute two experimental diets containing either 10(9) or 10(11) colony forming unit of live bacteria/g of feed while a third diet without the bacterial supplement served as the control diet. The diets were offered to rainbow trout (75g average weight) in triplicate tanks for 30 days. Fish were sampled at 10, 20 and 30 days after commencement of the feeding trial to determine the proportion of the given probiont in the gut microflora composition and the nonspecific humoral and cellular immune responses on the 30th day. The relative proportion of the probiont increased with the feeding duration in the intestine, but not in the stomach. The proportion of L. rhamnosus in the stomach corresponded to the intake levels while no such relation existed in the intestine. The serum lysozyme and complement activities were significantly greater in fish fed the higher level of probiont compared with the control fish. The phagocytic activity of head kidney leucocytes also showed similar tendencies. These observations indicate the potential immuno-regulatory role of probiotic organisms in rainbow trout.

Animal Nutritional Physiological Phenomena↗

Cytokine expression in leucocytes and gut cells of rainbow trout, Oncorhynchus mykiss Walbaum, induced by probiotics.

Understanding how the various host cells respond to probiotic bacteria in vitro may provide important insight into elaborate immune responses triggered by beneficial bacteria. The aim of this study was to investigate the detailed pattern of the mRNA expression of cytokines (IL-1beta, IL-8, TNF-alpha and TGF-beta) in head kidney (HK) leucocytes and gut cells isolated from rainbow trout (Oncorhynchus mykiss Walbaum) after co-culturing with live probiotics. HK leucocytes and gut cells adjusted to 5 x 10(6) and 2 x 10(6) ml(-1), respectively, in L-15 medium containing 25% decomplemented FCS and 300 mg l(-1) L-glutamine were co-cultured with Carnobacterium maltaromaticum B26 and C. divergens B33 at an multiplicity of infection of 25 for 6 and 12 h. Quantitative real-time reverse transcriptase polymerase chain reaction using SYBR Green I was employed to determine the mRNA expression of studied genes. Although neither probiotic strains significantly induced mRNA of the cytokines in gut cells, expression ratios of IL-1beta and TNF-alpha of HK cells were significantly higher, suggesting that these bacteria can stimulate innate immunity in rainbow trout.

Animals↗

Probiotics and prebiotics: role in clinical disease states.

Parents of pediatric patients are seeking alternatives to conventional therapy in the prevention and treatment of gastrointestinal disease states because of therapeutic failures caused by the increased incidence of antibiotic resistance. One such alternative is the use of probiotics and prebiotics to stimulate health-promoting indigenous flora to affect pathogen colonization and expression of disease. Probiotics are live flora given in oral quantities that allow for colonization of the colon. Probiotics are given as functional foods or dietary supplements, and function to activate the mucosal immune system and prevent pathogen colonization and translocation by strengthening the mucosal barrier, interfering with pathogen colonization, and in some instances, producing secretory antibacterial substances. Prebiotics are nondigestible carbohydrates, principally oligosoccharides, that are fermented by colonic commensals, stimulating their proliferation and producing short-chain fatty acids. Both protective nutrients have been shown to reduce the incidence and severity of infantile diarrhea, particularly rotaviral gastroenteritis, prevent antibiotic-induced diarrhea, and prevent and treat intestinal food allergy. With additional multicenter clinical trial confirmations, these substances may become routine in the care of infants and young children.

Child↗

Probiotics for animal nutrition in the European Union. Regulation and safety assessment.

Probiotics are alive micro-organisms, generally bacteria but also yeasts than, when ingested alive in sufficient amount, they have a positive effect on the health going beyond the nutritional ones commonly known. Probiotics may operate through a nutritional and/or health or sanitary effect. Micro-organisms used in animal feed in the EU are mainly bacterial strains of Gram-positive bacteria belonging to the types Bacillus, Enterococcus, Lactobacillus, Pediococcus, Streptococcus and strains of yeast belonging to the Saccharomyces cerevisiae species and kluyveromyces. While most of the species and genera are apparently safe, certain micro-organisms may be problematic, particularly the enterococci, which may harbour transmissible antibiotic resistance determinants and bacilli, specially those belonging to the Bacillus cereus group that are known to produce enterotoxins and an emetic toxin. The history and the current legislation in the European Union on probiotics feed additives including the requirements for the safety assessment for the target animal species, consumers, workers, and environment are presented.

Animal Feed↗

Probiotics and gastrointestinal health.

Evidence for positive health benefits of Lactobacilli applies to only a few strains used for commercial applications. It is generally agreed that a probiotic must be capable of colonizing the intestinal tract to influence human health; this requirement disqualifies many of the strains currently used in fermented dairy products. Lactobacillus GG, a variant of L. casei sps rhamnosus, has been studied extensively in adults and children. When consumed as a dairy product or as a lyophilized powder, LGG colonizes the gastrointestinal tract for 1-3 days in most individuals and up to 7 days in about 30% of subjects. Traveler's diarrhea, antibiotic-associated diarrhea, and relapsing Clostridium difficile colitis are improved with LGG. In infantile diarrhea, the severity and duration of the attack is reduced. LGG-fermented milk lessens the intestinal permeability defects caused by exposure to cows milk or rotavirus infection. LGG has proven beneficial effects on intestinal immunity. It increases the numbers of IgA and other immunoglobulin-secreting cells in the intestinal mucosa. LGG stimulates local release of interferon. It facilitates antigen transport to underlying lymphoid cells, which serves to increase antigen uptake in Peyer's patches. LGG also acts as an immunoadjuvant for oral vaccines. In an animal model of colon cancer, LGG reduced the incidence of chemically induced tumors in the large bowel of rodents. Extensive safety testing has shown no pathogenic potential in humans or animals. Probiotic cultures of Lactobacilli have the potential to bring substantial health benefits to the consumer. The purported benefits for any probiotic must pass the highest standards of scientific scrutiny before the claims can be accepted.

Adult↗

Probiotics and infectious diarrhea.

Numerous probiotic agents have been studied for the management of diarrheal disease. In particular, the prevention and management of acute viral diarrhea, the treatment of recurrent Clostridium difficile diarrhea, as well as the control of antibiotic-associated diarrhea seem to be areas of significant potential benefit. A few agents, including Lactobacillus GG, Lactobacillus reuteri, and Saccharomyces boulardii, seem to be promising agents for the amelioration of the course of acute diarrhea in children when used therapeutically. The use of other agents, particularly Bifidobacteria, supplementing the regular feed of infants may have an effect as prophylactic agents against acute diarrheal diseases. In general, in pediatric populations, the effect of probiotic agents appears to be most significant against viral (rotaviral) diarrhea, suggesting that an immunological mechanism is responsible for the beneficial effects. The numerous agents, doses, and populations used make generalization difficult. Nevertheless, it is clear probiotic agents are becoming an important part of the armamentarium against gastrointestinal problems in infants and children.

Child, Preschool↗

Probiotics and inflammatory bowel diseases.

The pathogenesis of inflammatory bowel diseases remains elusive. However, the resident luminal bacteria seem to be an important factor in their development and chronicity. There is evidence to suggest that inflammatory bowel diseases may represent an aggressive immunological response to the resident luminal flora, rather than an alteration in the normal flora. In prior research, probiotic bacteria were effective in managing certain acute diarrheal diseases, and investigators reported that certain Lactobacilli strains seem to have protective immunomodulating and bowel flora manipulating properties. We report the results of recent studies with probiotics in animal models, in which promising effects for the treatment of chronic inflammatory bowel disease, pouchitis, and ulcerative colitis were observed. Future research may clarify a precise role for probiotic bacteria in managing chronic inflammatory bowel diseases.

Animals↗

Antimutagenic properties of probiotic bacteria and of organic acids.

Antimutagenic activities of live and killed cells of 6 strains of Lactobacillus acidophilus and 9 strains of bifidobacteria and of organic acids usually produced by these probiotic bacteria were determined using 8 potent chemical mutagens and promutagens. The mutagens and promutagens used were N-methyl, N'-nitro, N-nitrosoguanidine; 2-nitroflourene; 4-nitro-O-phenylenediamine; 4-nitroquinoline-N-oxide; Aflatoxin-B; 2-amino-3-methyl-3H-imidazoquinoline; 2-amino-1-methyl-6-phenyl-imidazo (4,5-b) pyridine, and 2-amino-3-methyl-9H-pyrido (3,3-6) indole. The mutagenicity of these mutagens and antimutagenic activity of probiotic bacteria against the mutagens were determined according to the Ames TA-100 assay using a mutant of Salmonella typhimurium. Efficiency of bacterial cells in binding or inhibiting these mutagens was also investigated. Live cells of probiotic bacteria showed higher antimutagenic activity and their efficiency in inhibiting the mutagens was better than killed bacterial cells. Live bacterial cells bound or inhibited the mutagens permanently, whereas killed bacteria released mutagens upon extraction with dimethyl sulfoxide. Among the organic acids, butyric acid showed highest inhibition of mutagens followed by acetic acid. Lactic and pyruvic acids did not show appreciable levels of inhibition.

Bifidobacterium↗

Probiotics in primary prevention of atopic disease: a randomised placebo-controlled trial.

BACKGROUND: Reversal of the progressive increase in frequency of atopic disease would be an important breakthrough for health care and wellbeing in western societies. In the hygiene hypothesis this increase is attributed to reduced microbial exposure in early life. Probiotics are cultures of potentially beneficial bacteria of the healthy gut microflora. We assessed the effect on atopic disease of Lactobacillus GG (which is safe at an early age and effective in treatment of allergic inflammation and food allergy). METHODS: In a double-blind, randomised placebo-controlled trial we gave Lactobacillus GG prenatally to mothers who had at least one first-degree relative (or partner) with atopic eczema, allergic rhinitis, or asthma, and postnatally for 6 months to their infants. Chronic recurring atopic eczema, which is the main sign of atopic disease in the first years of life, was the primary endpoint. FINDINGS: Atopic eczema was diagnosed in 46 of 132 (35%) children aged 2 years. Asthma was diagnosed in six of these children and allergic rhinitis in one. The frequency of atopic eczema in the probiotic group was half that of the placebo group (15/64 [23%] vs 31/68 [46%]; relative risk 0.51 [95% CI 0.32-0.84]). The number needed to treat was 4.5 (95% CI 2.6-15.6). INTERPRETATIONS: Lactobacillus GG was effective in prevention of early atopic disease in children at high risk. Thus, gut microflora might be a hitherto unexplored source of natural immunomodulators and probiotics, for prevention of atopic disease.

Child, Preschool↗

Potential probiotic lactic acid bacteria Lactobacillus rhamnosus (HN001), Lactobacillus acidophilus (HN017) and Bifidobacterium lactis (HN019) do not degrade gastric mucin in vitro.

The mucus layer (mucin) coating the surface of the gastrointestinal tract (GIT) plays an important role in the mucosal barrier system. Any damage or disturbance of this mucin layer will compromise the host's mucosal defence function. In the present study, the ability of three potential probiotic lactic acid bacteria (LAB) strains (Lactobacillus rhamnosus HN001, Lactobacillus acidophilus HN017, Bifidobacterium lactis HN019) to degrade mucin in vitro was evaluated, in order to assess their potential pathogenicity and local toxicity. The LAB strains were incubated in medium containing hog gastric mucin (HGM, 0.3%) at 37 degrees C for 48 h, following which any decrease in carbohydrate and protein concentration in the ethanol-precipitated portion of the culture medium was determined, using phenol-sulphuric acid and bicinchonic acid (BCA) protein assays, respectively. The change in molecular weight of mucin glycoproteins, following incubation with the test strains, was monitored by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). In order to expose any ability of the test strains to degrade mucin visually and more directly, the test strains were also cultured on agarose containing 0.3% HGM and incubated anaerobically for 72 h at 37 degrees C. No significant change in the carbohydrate or protein concentration in mucin substrates was found following incubation with the test strains. No mucin fragments were derived from the mucin suspension incubated with test strains, and no mucinolysis zone was identified on agarose. These results demonstrate that the potential probiotic LAB strains tested here were unable to degrade gastrointestinal mucin in vitro, which suggests that these novel probiotic candidates are likely to be non-invasive and non-toxic at the mucosal interface.

Anaerobiosis↗

Safety and efficacy of probiotic lactobacilli in promoting growth in post-weaning Swiss mice.

Lactobacillus rhamnosus MA27/6B and L. acidophilus MA27/6R are strains used in feed as probiotics. Their safety profiles and growth-stimulating properties were investigated via in vivo studies on young Swiss mice. After repeated administrations of different probiotic preparations in drinking water, safety parameters determined from liver, spleen and total weight remained unchanged. The growth-stimulating properties of viable or dead lactobacilli were studied after supplementation in drinking water. The feed intake (FI), water intake and body weight gain (WG) of the animals were compared to those of control mice. The lactobacilli supplementation of a sub-optimal diet made of barley allow recording of measurable growth performance of mice. It significantly increased WG compared to control groups (P < 0.01), by +28.9% and +31.7% for L. rhamnosus MA27/6B and L. acidophilus MA27/6R, respectively. This WG was correlated with a decrease in the consumption index. The effect of the dose ingested was also investigated: 10(8) lactobacilli CFU/mouse/day produced greater WG than 10(2), 10(4) or 10(6) CFU/mouse/day. No significant differences in growth performance parameters were observed between mice fed with 10(8) cells of viable or nonviable preparations. The mouse assay described could be used as a preliminary criterion when screening candidate probiotics for growth performance properties.

Animals↗

Effect of a probiotic formula on intestinal immunoglobulin A production in healthy children.

The anti-infectious effect of probiotics has recently been reported and one mechanism may be the non-specific stimulation of immunity. This study was performed to elucidate the influence of a probiotic formula on intestinal microflora and local immunity in healthy children. A follow-up formula containing viable bifidobacteria was given to seven healthy Japanese children (15 to 31 months old) for 21 days. During intake of the formula, the administered strain was detected in feces from five subjects (71%) and total fecal bifidobacteria slightly increased. Fecal levels of total IgA and anti-poliovirus IgA during intake of the formula were significantly higher than those before intake (P < 0.05). The increase in local IgA levels resulting from ingestion of the probiotic formula may contribute to enhancement of the mucosal resistance against gastrointestinal infections.

Antibodies, Viral↗