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Probiotics and prebiotics in chronic inflammatory bowel diseases.

The prokaryotic and eukaryotic cells of the colon exist in a highly complex, but harmonious relationship. Disturbances in this remarkable symbiosis can result in the development of inflammatory bowel diseases (IBD). Although the etiology of IBD is not entirely understood, it is known that the chronic inflammation of Crohn's disease, ulcerative colitis and chronic pouchitis are a result of an overly aggressive immune response to the commensal intestinal flora in genetically susceptible hosts. Recent studies have enhanced our ability to understand the interaction between the host and its intestinal microflora and the role the microflora plays in maintaining intestinal homeostasis. As we begin to understand the benefits conferred to the intestine by the microflora, the notion of modifying the composition of the bacterial load to improve human health has arisen. A significant body of research now exists investigating the role of probiotics and prebiotics in ameliorating chronic intestinal inflammation. This article will begin with an overview of the role of the commensal microflora in maintaining mucosal immune homeostasis, and how a dysregulated immune response to the intestinal microflora results in IBD. This will be followed by a summary of the use of probiotics and prebiotics in experimental and human IBD.

Animals↗

Lactobacilli isolated from chicken intestines: potential use as probiotics.

Lactobacillus strains were tested for their in vitro probiotic properties. Cell surface hydrophobicity was found to be very high for Lactobacillus fermentum subsp. cellobiosus and Salmonella Gallinarum; high values could indicate a greater ability to adhere to epithelial cells. Studies on Lactobacillus animalis indicated relative cell surface hydrophobicities smaller than those of L. fermentum subsp. cellobiosus and L. fermentum. L. animalis and Enterococcus faecalis were able to coaggregate with L. fermentum subsp. cellobiosus and L. fermentum, respectively, but not with Salmonella Gallinarum. After mixed-culture studies for determining suitable growth behavior, the pair of strains L. animalis plus L. fermentum subsp. cellobiosus was selected for an attempted challenge against Salmonella Gallinarum. Double and triple mixed-culture studies indicated that selected lactobacillus strains were able to retain their beneficial characteristics in the presence of Salmonella Gallinarum such as presence of lectins, production of antimicrobial compounds, and ability to grow and compete. The selected microorganisms can be considered as potential ingredients for a chicken probiotic feed formulation intended to control salmonellosis and also improve poultry sanitation.

Animals↗

Inability of probiotic bacterial strains Lactobacillus rhamnosus HN001 and Bifidobacterium lactis HN019 to induce human platelet aggregation in vitro.

Platelet aggregation contributes to the pathogenesis of infective endocarditis, and aggregation of platelets induced by lactobacilli is thought to be an important contributory factor in the development and progression of Lactobacillus endocarditis. The main purpose of this study was to examine the effect of immunity-enhancing probiotic strains Lactobacillus rhamnosus HN001 and Bifidobacterium lactis HN019 on the activation and aggregation of human blood platelets. Whole blood samples from healthy individuals were incubated in vitro with HN001 or HN019 and subsequently labeled with platelet-specific monoclonal antibodies, fluorescein isothiocyanate-conjugated anti-CD41a (expressed on normal platelets), and phycoerythrin-streptavidin-conjugated anti-CD62p (expressed on activated platelets) before analysis by flow cytometry. Platelet-rich plasma was used to assist the gating of the platelet cluster. ADP and epinephrine were used as the physiological platelet activation agonists. Platelet aggregation-inducing strain Streptococcus sanguis 133-79 was used as a positive control strain. The mean fluorescence intensity of phycoerythrin and the percentage of platelets expressing the CD62p marker were used to assess the degree of platelet activation. The percentage of CD62p-positive platelets and the light scatter profiles of the agonist-activated platelets were used to identify the occurrence and degree of platelet aggregation. HN001 and HN019 had no effect on spontaneous platelet activation and aggregation; they also failed to exacerbate the platelet aggregation activity induced by ADP and epinephrine. Therefore, these test probiotic strains HN001 and HN019 are less likely to participate in the pathogenesis of infective endocarditis or other thrombotic disorders with regard to platelet aggregation factors.

Antibodies, Monoclonal↗

[The antibacterial resistance of the strains making up the components of probiotic preparations].

The spectrum of the antibiotic resistance of 21 strains of normal microflora, mainly forming constituents of widely used probiotics to 25 antibacterial preparations. Lactobacillus spp. were prevalent in the strains under test. The spectrum of the antibiotic resistance of lactobacilli varied, the gradation of resistance being more pronounced with respect to strains and not species. Both highly sensitive and highly resistant Lactobacillus strains were found: L. acidophilus (a component of biopreparation "Linex"), L. plantarum 8RA3, L. fermentum 90T4C (probiotic "Lactobacterin"), L. fermentum BL96. Bacteria used as the components of combined preparation "Linex" exhibited the highest resistance to a number of modern antibiotics. Strains of bifidobacteria were found to be highly sensitive to antibiotics.

Bifidobacterium↗

[Probiotics: Is there a scientific basis for their effects?].

Probiotic microorganisms have been used with increasing frequency in nutrition and for therapeutic purposes during the last years. Their application is generally based upon empirical experiences, since their mode of action has been the topic of investigations only recently. It became apparent that probiotics exert a wide spectrum of different effects ranging from direct antagonism against pathogens to influence upon intestinal epithelium and immune system of the host's organism. In this review, the current knowledge is summarized.

Digestive System↗

Could probiotics be an option for treating and preventing urogenital infections?

Considering the enormity of the problem in terms of women infected per year, urogenital infections receive far too little attention from scientists, government funding agencies, and the pharmaceutical industry. A recent resurgence in interest among clinicians is a result of consumer demands for better therapies, problems resulting from drug resistance, and the prospect of new diagnostics and treatments on the horizon. It is now recognized that the intestinal and urogenital microflora are critical for the health and well-being of humans. The concept of replenishing these flora with probiotic organisms seems to be an option that has a growing scientific basis. Although few strains have been selected and targeted for urogenital applications, and none are currently available on the market, evidence shows that probiotic therapy has the potential to make an impact on women's health.

Canada↗

Anti-carcinogenicity of probiotics and prebiotics.

Yoghurt, and the lactic acid producing bacteria (LAB; probiotics) that it contains, have received much attention as potential cancer-preventing agents in the diet. It is usually considered that the mechanism of the action is by increasing the numbers of LAB in the colon, which modifies the ability of the microflora to produce carcinogens. Prebiotics such as non-digestible oligosaccharides (NDO) appear to have similar effects on the microflora by selectively stimulating the growth of LAB in the colon. Evidence for cancer-preventing properties of pro- and prebiotics is derived from studies on faecal enzyme activities in animals and humans, inhibition of genotoxicity of known carcinogens in vitro and In vivo, suppression of carcinogen-induced preneoplastic lesions and tumours in laboratory animals. Some of these studies indicate that combinations of pro and prebiotics ('synbiotics') are more effective. Epidemiological and intervention studies provide some, albelt limited, evidence for protective effects of products containing probiotics in humans.

Animals↗

The development of gut immune responses and gut microbiota: effects of probiotics in prevention and treatment of allergic disease.

The infant's immature intestinal immune system develops as it comes into contact with dietary and microbial antigens in the gut. The evolving indigenous intestinal microbiota have a significant impact on the developing immune system and there is accumulating evidence indicating that an intimate interaction between gut microbiota and host defence mechanisms is mandatory for the development and maintenance of a balance between tolerance to innocuous antigens and capability of mounting an inflammatory response towards potential pathogens. Disturbances in the mucosal immune system are reflected in the composition of the gut microbiota and vice versa. Distinctive alterations in the composition of the gut microbiota appear to precede the manifestation of atopic disease, which suggests a role for the interaction between the intestinal immune system and specific strains of the microbiota in the pathogenesis of allergic disorders. The administration of probiotics, strains of bacteria from the healthy human gut microbiota, have been shown to stimulate antiinflammatory, tolerogenic immune responses, the lack of which has been implied in the development of atopic disorders. Thus probiotics may prove beneficial in the prevention and alleviation of allergic disease.

Food Hypersensitivity↗

[The probiotic Lactobacillus acidophilus--an alternative treatment of bacterial vaginosis].

The probiotics or biotherapeutical agents are microorganisms that have an antagonistic activity regarding pathogens in vivo. Therapeutic application of these microorganisms is mostly in cases of infections of non-sterile mucous surfaces--vagina and large intestines. With the help of the probiotic L. acidophilus bacterial vaginosis can be treated successfully in a natural way. The lack of systemic side effects makes it a drug of choice in the treatment of pregnant women. More studies in this direction and development of new strategies for treatment of bacterial vaginosis in pregnant and non-pregnant women are needed.

Clinical Trials as Topic↗

Bacteriological evaluation of dog and cat diets that claim to contain probiotics.

Nineteen commercial pet foods claiming to contain probiotics were evaluated. Selective bacterial culture was performed to identify organisms that were claimed to be present. Twelve diets claimed only to contain specific bacterial fermentation products, which does not necessarily indicate that live growth would be expected, but these products also included the term "probiotic" somewhere on the package, suggesting that live, beneficial organisms were present. No products contained all of the listed organisms, while 1 or more of the listed contents were isolated from 10 out of 19 products (53%). Eleven products contained additional, related organisms including Pediococcus spp, which was isolated from 4 products. No relevant growth was present in 5 (26%) products. Average bacterial growth ranged from 0 to 1.8 x 10(5) CFU/g. Overall, the actual contents of the diets were not accurately represented by the label descriptions.

Animal Feed↗

Effect of fermented milk containing the probiotic Lactobacillus casei DN-114001 on winter infections in free-living elderly subjects: a randomised, controlled pilot study.

Probiotics are being increasingly studied for their ability to enhance host resistance to, and recovery from, infection. The probiotic strain Lactobacillus casei DN-114001 has previously been shown to reduce the incidence and duration of episodes of diarrhoea in children. Our controlled pilot study aimed to evaluate the effect of supplementation for 3 weeks with milk fermented with yoghurt cultures and L. casei DN-114001 on the incidence and severity of winter infections (gastrointestinal and respiratory) in elderly people. We found no difference in the incidence of winter infections between groups. However, duration of all pathologies was significantly lower in the treatment group (7.0 3.2 days, n=180) than in the control group (8.7 3.7 days; n=180) (p=0.024), as was maximal temperature (38.3 0.5 C treatment group vs. 38.5 0.6 C control; p=0.01). The potential for a 20% reduction in the duration of winter infections that we have found warrants further investigation on a larger scale.

Aging↗

Preliminary investigation of the probiotic potential of Lactobacillus rhamnosus strain GG in horses: fecal recovery following oral administration and safety.

This study was designed to evaluate whether Lactobacillus rhamnosus strain GG (LGG), an extensively studied probiotic organism in humans, can colonize the intestines of adult horses and foals. Lactobacillus rhamnosus strain GG was administered to adult horses at doses of 1 x 10(9) CFU/50kg bodyweight (BW)/day (group 1, 7 horses), 1 x 10(10) colony forming units/ 50kg BW/day (group 2, 7 horses) and 5 x 10(10) colony forming units/50kg BW/day (group 3, 7 horses) for 5 d. Foals received 2 x 10(10) colony forming units/50kg BW/day (group 1, 7 foals) or 1 x 10(11) colony forming units/50kg BW/day (group 2, 7 foals) for 5 d. Fecal levels of L. rhamnosus strain GG in adult horses were low and variable in the 2 lower dose groups. Even in the high dose group, colonization was relatively low. In contrast, more consistent intestinal colonization was present in foals, and colonization persisted for up to 9 d following cessation of administration. No adverse effects were observed in any animal. Clinical studies evaluating this probiotic are indicated in foals. The presence of this organism in the feces of adult horses may only represent passive movement through the intestinal tract, not actual colonization. Consistent intestinal colonization in adults was only achieved with a prohibitively high dose.

Administration, Oral↗

Assessment of mucosal inflammation and circulation in response to probiotics in patients operated with ileal pouch anal anastomosis for ulcerative colitis.

BACKGROUND: Pouchitis is a common and troublesome condition, and a disturbed microbiological flora and mucosal blood flow in the pouch have been suggested as possible causes. Laser Doppler flowmetry (LDF) has been used successfully to measure gastric and colonic mucosal perfusion in humans. The aim of this study was to evaluate the effect of intervention with probiotics on ileal pouch inflammation and perfusion in the pouch, assessed by endoscopy, histology, fecal calprotectin and LDF. METHODS: A fermented milk product (Cultura; 500 ml) containing live lactobacilli (La-5) and bifidobacteria (Bb-12) was given daily for 4 weeks to 10 patients operated with ileal-pouch-anal anastomosis (IPAA) for ulcerative colitis (UC). Mucosal perfusion was measured with LDF and the degree of inflammation was examined at predefined levels of the distal bowel by endoscopy and histology. Stool samples were cultured for lactobacilli and bifidobacteria and calprotectin were measured before and after intervention. RESULTS: The LDF measurements were reproducible in the pelvic pouch at each of the predefined levels, but did not change after intervention. The mucosal perfusion was reduced in the distal compared to the proximal part of the pouch. Calprotectin levels did not change significantly after intervention. The median endoscopic score for inflammation was significantly reduced by 50% after intervention, whereas the histological score did not change significantly. CONCLUSION: The results suggest that probiotics primarily act superficially, with change of gross appearance of the mucosa at endoscopy, but without significant effect on histological picture, mucosal perfusion or faecal calprotectin, during a relatively short period of 4 weeks.

Adolescent↗

Enumeration of probiotic bacilli spores in animal feed: interlaboratory study.

Fourteen out of 17 laboratories completed an interlaboratory study comparing 2 pretreatment protocols of feed samples containing authorized probiotic bacilli spores. Both methods used tryptone soy agar for enumeration. Pretreatment A involved preparation of a suspension of the feed sample in 50% ethanol. For pretreatment B, the sample was suspended in peptone salt solution and heated at 80 degrees C for 10 min. Each laboratory analyzed 12 samples (6 per pretreatment), which represented duplicates of a high (10(9) colony-forming units [CFU]/g) and low (10(5) CFU/g) level of bacilli spores or a blank that contained vegetative probiotic bacteria only. For pretreatment A, the repeatability relative standard deviation (RSD(r)) was 2.9% for the low level and 2.5% for the high. The reproducibility relative standard deviation (RSDR) values were 7.8 and 5.9%, respectively. Pretreatment B revealed RSD(r) values of 1.1 and 1.0%, and RSDR values of 5.8 and 3.4%, respectively. The heat treatment (pretreatment B) of feed samples had better precision data, resulted in higher viable bacilli counts, and was more effective in deactivating vegetative background flora. It is therefore recommended for adoption for official control purposes and for CEN and ISO standards.

Animal Feed↗

[Biological effects of interferon, produced by recombinant bacteria of the probiotic preparation subalin].

The present review deals with the analysis of biological and functional activities of recombinant bacteria Bacillus subtilis IF-alpha 2335 are producing a human interferon. The interferon-producing bacteria are constructed on a basis commercial probiotic strain B.subtilis 2335, carrying a recombinant plasmid pMBM 105 with the gene of human alpha-2 interferon. The implementation of the recombinant strain in the preparation probiotic, received a designation "Subalin", necessitates to verify a number of immunologic activities and to perform successive protective effects. Interferon, synthesized by recombinant bacteria shows the activity on macroorganism at oral and rectal application of preparation. Subalin was shown antivirus and antitumor activity and preservation by recombinat bacteria of antagonistic properties. The mechanisms of the positive effect of subalin were considered: this effect was shown to be due to the action of interferon excreted by recombinant bacteria into the mucous of different biotopes of host.

Administration, Oral↗

[Spore probiotics].

Data on probiotic preparations prepared from spore-forming bacteria is reviewed. Information on indications for use, effectiveness and the mechanism of action is given. Preparations being produced in Russia (Biosporin, Sporobacterin and Bactosporin) are evaluated in detail as well as the prospects of the application of probiotics from spore-forming bacteria.

Animals↗

[Use of molecular methods in food microbiology with the example of probiotic use of lactobacilli].

The aim of molecular methods in food microbiology is the identification or strain specific differentiation of microorganisms. Identification methods include besides taxonomic purposes also the detection of virulence genes or resistance markers. Strain specific differentiation is used for epidemiological investigations or the quality control of technologically used bacteria. Probiotic strains of the Lactobacillus acidophilus-group were investigated with different molecular methods: for identification proteinfingerprinting and RAPD-PCR (Random Amplified Polymorphic DNA-PCR) were applied, for strain specific differentiation pulsed field gel electrophoresis (PFGE) and again RAPD-PCR were used. The molecular methods applied should be chosen depending on the objective (identification, differentiation) and with respect to the organism to be tested. In case of probiotic lactic acid bacteria like the L. acidophilus-group proteinfingerprinting has proved to be successful for identification and with some limitation also RAPD-PCR. For differentiation PFGE is suitable as well as RAPD-PCR. Those methods differ substantially in their work load and in personal requirements, and show different power of discrimination and reproducibility within and between laboratories. This should be considered while choosing the appropriate method.

DNA, Bacterial↗

Chemopreventive effect of a probiotic preparation on the development of preneoplastic and neoplastic colonic lesions: an experimental study.

BACKGROUND/AIMS: A number of studies have suggested a key role played by certain resident gut bacteria in the development of large bowel cancer. The aim of the present study was to test the effect of a novel symbiotic preparation, which has been recently shown to beneficially modify gut ecosystem and systemic immunity, on either preneoplastic and neoplastic changes in a colon carcinogenesis model. METHODOLOGY: Sprague-Dawley rats were fed a standard diet for 1 week and then were randomly assigned to three groups. The control diet was given to groups A and B, whereas in group C, the same diet plus 2 mL of a probiotic mixture was given throughout the experiment. Thirty rats (groups B, C) each received a weekly subcutaneous injection of azoxymethane at a dose of 15 mg/kg of body weight for 10 weeks. Group A served as a control group and received a subcutaneous injection of saline for 10 weeks. Forty-five rats were sacrificed at 3-week observation and 60 rats at 20-week observation for assessing metaphase index together with aberrant crypt foci and intestinal immune system markers from one hand and tumor occurrence from the other, respectively. RESULTS: Group A showed a significantly increased metaphase index either in aberrant crypt foci or in "normal appearing" crypts when compared to group A (p < 0.01). Group B rats caused a significant decrease at both sites (p < 0.05). The numbers of lymphocytes derived from the mesenteric lymph nodes in group B rats were significantly decreased (p < 0.01) as compared to either control and to group C. The percentage of CD8 lymphocytes in group C was significantly higher than that in group B. Group C showed a significantly reduced ratio of aberrant crypt foci/colon and of aberrant crypt per colon and per each single focus (p < 0.05). A total of 18 (90%) group B and 10 (50%) group C rats had colon tumors, this difference was significant. The mean number of colon tumors per rat was 2.2 and 1.0 in group B and C, respectively. CONCLUSIONS: Effective probiotics treatment, through mechanisms still to be fully elucidated (decreased fecal pH, specific reduction of carcinogenetic bacterial enzymes, modulation of gut-associated and systemic immune system etc.) has the potential to exert significant antimutagenic properties against colon cancer.

Animals↗