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Probiotics and inflammatory bowel disease.

Inflammatory bowel disease (IBD) is characterised by a chronic dysregulation of the inflammatory response in the gastrointestinal tract. While the pathogenesis is unclear, studies have demonstrated that the gastrointestinal tracts of patients with IBD are populated with increased levels of adherent and pathogenic bacteria. This evidence, combined with growing data accumulated from genetic studies as well as animal models of IBD, indicates that an aberrant response to altered enteric flora plays a significant role in the disease process. Current therapies for IBD have been directed towards the development of anti-inflammatory agents and immunomodulators to attenuate the inflammatory response in the gastrointestinal tract. Antibiotics are also partially effective in the treatment of IBD, presumably by altering the bowel flora. However, it is clear from clinical trials that immunomodulators and antibiotics are not effective in a large proportion of patients with IBD and other therapeutic alternatives need to be pursued. Probiotics are microbial supplements capable of recolonising the bowel with non-pathogenic strains of bacteria or yeast. Probiotics have long been shown to be beneficial in both infectious and non-infectious digestive disorders. Growing evidence indicates that probiotics may be effective in the treatment of specific clinical IBD conditions. This article addresses the current evidence for the role of enteric flora in the pathogenesis of IBD and the clinical evidence supporting the use of probiotics in specific clinical IBD conditions.

Animals↗

Probiotics as biotherapeutic agents: present knowledge and future prospects.

Since the early observations of Elie Metchnikoff, a wealth of experiments have described the use of selected microorganisms, mainly belonging to the lactic acid bacteria family, for the prevention or treatment of a variety of pathological situations. Nevertheless, the mechanisms underlying the proposed actions remain vastly unknown, partly as a consequence of the complexity of the gastro-intestinal ecosystem with which these biotherapeutic agents are expected to interact, but also because of the increasing variety of strains considered to have potential probiotic characteristics. During the past decades, however, the beneficial effect of specific strains in preventing or treating intestinal disorders has been substantiated by well-controlled clinical trials. Increasing evidence, including human studies, is also supporting the immunomodulatory role attributed to given lactic acid bacterial strains. The desire by consumers to use natural methods for health maintenance rather than long-term chemotherapeutic agents (i.e. antibiotics), linked to their expectation that food becomes a source of prolonged well-being, supports the speculation that the probiotic market will expand rapidly. Much of this growth will also depend on the reliability of claims that these products will bare. Therefore, the legislator will have to provide clear rules and regulations which will depend on measurable biomarkers and criteria based on scientific evidence. These commercial and legislative needs will hopefully provide scientists with the resources necessary to conduct the multidisciplinary research required to establish facts and mechanisms of action for carefully selected probiotic strains. These research results will probably be as essential for the positioning of probiotic preparations as either a food, a food supplement or as pharmaceutical preparation.

Animals↗

Probiotics: an emerging therapy.

There is considerable clinical interest in the utility of probiotic therapy--the feeding of (live) non-pathogenic bacteria, originally derived from the alimentary tract, for disease treatment or health promotion. The microflora of the gastrointestinal tract is essential for mucosal protection, for immune education and for metabolism of fecal residue. Physiological disturbances of these processes, when they occur, result from: i) alteration of a microbial ecosystem, originally conserved by evolution; ii) reduced consumption of microorganisms; iii) invasion of pathogens; or iv) modern interventions. Recent data support the use of proven probiotic organisms in prevention and treatment of flora-related gastrointestinal disorders including inflammatory bowel disease, infectious and antibiotic related diarrheas, and post-resection disorders including pouchitis. Therapeutic activity of probiotic bacteria can be due to competition with pathogens for nutrients and mucosal adherence, production of antimicrobial substances, and modulation of mucosal immune functions. Although a promising treatment, controlled clinical trials are necessary to validate the benefit of probiotics.

Animals↗

Screening of yeasts as probiotic based on capacities to colonize the gastrointestinal tract and to protect against enteropathogen challenge in mice.

Probiotics are defined as viable microorganisms that exhibit a beneficial effect on the host's health when they are ingested. Two important criteria are used for selection of probiotic microorganisms: they must be able to survive in the gastrointestinal environment and to present at least one beneficial function (colonization resistance, immunomodulation or nutritional contribution). Generally, in vitro assays demonstrating these properties were used to select probiotics but it is unclear if the data can be extrapolated to in vivo conditions. In the present work, twelve Saccharomyces cerevisiae strains isolated from different environments (insect association, tropical fruit, cheese and "aguardente" production) and pre-selected for in vitro resistance to simulated gastrointestinal conditions were inoculated in germ-free mice to evaluate their real capacity to colonize the mammal digestive tract. Using these data, one of the yeasts (S. cerevisiae 905) was selected and tested in gnotobiotic (GN) and conventional (CV) mice for its capacity to protect against oral challenge with two enteropathogenic bacteria (Salmonella Typhimurium and Clostridium difficile). The yeast reached populational levels potentially functional in the gastrointestinal portions where the enteropathogens tested act. No antagonism against either pathogenic bacterium by the yeast was observed in the digestive tract of GN mice but, after challenge with S. Typhimurium, mortality was lower and liver tissue was better preserved in CV animals treated with the yeast when compared with a control group (p<0.05). Histopathological results of intestines showed that the yeast also presented a good protective effect against oral challenge with C. difficile in GN mice (p<0.05). In conclusion, among the 12 S. cerevisiae tested, strain 905 showed the best characteristics to be used as a probiotic as demonstrated by survival capacity in the gastrointestinal tract and protective effect of animals during experimental infections.

Animals↗

Species-specific identification of commercial probiotic strains.

Products containing probiotic bacteria are gaining popularity, increasing the importance of their accurate speciation. Unfortunately, studies have suggested that improper labeling of probiotic species is common in commercial products. Species identification of a bank of commercial probiotic strains was attempted using partial 16S rDNA sequencing, carbohydrate fermentation analysis, and cellular fatty acid methyl ester analysis. Results from partial 16S rDNA sequencing indicated discrepancies between species designations for 26 out of 58 strains tested, including two ATCC Lactobacillus strains. When considering only the commercial strains obtained directly from the manufacturers, 14 of 29 strains carried species designations different from those obtained by partial 16S rDNA sequencing. Strains from six commercial products were species not listed on the label. The discrepancies mainly occurred in Lactobacillus acidophilus and Lactobacillus casei groups. Carbohydrate fermentation analysis was not sensitive enough to identify species within the L. acidophilus group. Fatty acid methyl ester analysis was found to be variable and inaccurate and is not recommended to identify probiotic lactobacilli.

Carbohydrate Metabolism↗

Metabolic and biochemical responses of probiotic bacteria to oxygen.

The interaction between oxygen and probiotic bacteria was studied by growing Lactobacillus acidophilus and Bifidobacterium spp. in 0, 5, 10, 15, and 21% oxygen in a hypoxic glove box. The metabolic responses of each probiotic strain in the different oxygen environments were monitored by measuring the levels of lactic acid and determining the lactate-to-acetate ratio. Biochemical changes induced by oxygen were examined by monitoring the specific activities of NADH oxidase, NADH peroxidase, and superoxide dismutase. In addition, the ability to decompose hydrogen peroxide and the sensitivity of each strain to hydrogen peroxide was also determined. With an increase in oxygen percentage, levels of lactic acid in L. acidophilus strains decreased, whereas the lactate-to-acetate ratio reduced in all the bifidobacteria tested. At 21% oxygen, the specific activities of NADH oxidase and NADH peroxidase, and the hydrogen peroxide decomposing ability of five probiotic strains was significantly higher than at 0% oxygen. The sensitivity of the probiotic strains to hydrogen peroxide however, remained unaffected in all the different oxygen percentages. Superoxide dismutase levels did not reveal any conclusive trend. In both L. acidophilus and Bifidobacterium spp., NADH oxidase and NADH peroxidase functioned optimally at pH 5. Growth in the various oxygen environments did not change this optimum pH.

Bifidobacterium↗

Probiotic Lacticaseibacillus casei 2S-1 Attenuates Escherichia coli-Induced Enteritis via Gut Microbiota Modulation and Host Gene Regulation.

Maintaining gut microbial homeostasis is crucial for host health, whereas infection with Escherichia coli (E. coli) is a major contributor to intestinal inflammation and microbial dysbiosis. Recent research has focused on probiotic strategies for managing enteric inflammatory disorders. Previous studies have shown that beneficial microorganisms show protection through modulating host immune responses, enhancing intestinal epithelial barrier integrity, and inhibiting pathogenic bacteria. To evaluate the prophylactic effectiveness of a recently isolated strain, Lacticaseibacillus casei 2S-1, in a murine model of E. coli-induced enteritis, this study focuses on interactions within the microbiota-intestinal-immune axis, together with host transcriptional responses and pathway enrichment associated with oxidative stress and mitochondrial function. In vitro analysis of probiotic features, including growth dynamics, acidogenic capacity, and tolerance to acidic and bile salt environments, as well as genetic safety profiling, followed the methodical isolation and taxonomic identification of L. casei 2S-1. A preventive intervention protocol was established, and a murine model of enteritis was induced by exposure to E. coli. Histopathological analyses were performed to observe in vivo safety and protective efficacy. Changes in gut microbial structure were characterized by 16S rRNA gene sequencing, while host responses were identified by intestinal immunohistochemistry and transcriptome profiling. L. casei 2S-1 showed probiotic properties. In vitro analyses showed that the strain exhibited tolerance to acidic and bile salt conditions, and its untreated culture supernatant showed antimicrobial activity against pathogenic bacteria. Its safety profile was supported by genomic analysis, which verified the lack of virulence-associated genes and antibiotic resistance factors. In vivo, L. casei 2S-1 pretreatment reduced mortality and intestinal inflammation, modulated gut microbial composition, and preserved intestinal barrier-associated protein expression in infected mice. This study provides experimental evidence supporting the prophylactic effects of L. casei 2S-1 and its associations with gut microbiota modulation and host transcriptional responses, providing a foundation for further investigation of probiotic-based preventive strategies against intestinal infections.

Animals↗

Effects of probiotic on intestinal mucosa of patients with ulcerative colitis.

AIM: To investigate the effects of probiotic on intestinal mucosae of patients with ulcerative colitis (UC), and to evaluate the role of probiotic in preventing the relapse of UC. METHODS: Thirty patients received treatment with sulphasalazine (SASP) and glucocorticoid and then were randomly administered bifid triple viable capsule (BIFICO) (1.26 g/d), or an identical placebo (starch) for 8 wk. Fecal samples were collected for stool culture 2 wk before and after the randomized treatments. The patients were evaluated clinically, endoscopically and histologically after 2 mo of treatment or in case of relapse of UC. p65 and IkappaB expressions were determined by Western blot analysis. DNA-binding activity of NF-kappaB in colonic nuclear extracts was detected by electrophoretic mobility shift assay (EMSA). mRNA expressions of cytokines were identified by semi-quantitative assay, reverse transcriptase- polymerase chain reaction (RT-PCR). RESULTS: Three patients (20%) in the BIFICO group had relapses during 2-mo follow-up period, compared with 14 (93.3%) in placebo group (P<0.01). The concentration of fecal lactobacilli, bifidobacteria was significantly increased in BIFICO-treated group only (P<0.01). The expressions of NF-kappaB p65 and DNA binding activity of NF-kappaB were significantly attenuated in the treatment group than that in control (P<0.05). The mRNA expression of anti-inflammatory cytokines was elevated in comparison with the control group. CONCLUSION: The probiotic could impede the activation of NF-kappaB, decrease the expressions of TNF-alpha and IL-1beta and elevate the expression of IL-10. These results suggest that oral administration of this new probiotic preparation is effective in preventing flare-ups of chronic UC. It may become a prophylactic drug to decrease the relapse of UC.

Anti-Inflammatory Agents, Non-Steroidal↗

Antibiotics and probiotics in treatment of inflammatory bowel disease.

Many experimental and clinical observations suggest that intestinal microflora plays a potential role in the pathogenesis of inflammatory bowel disease (IBD). Manipulation of the luminal content using antibiotics or probiotics represents a potentially effective therapeutic option. The available studies do not support the use of antibiotics in ulcerative colitis (UC). Antibiotics are effective in treating septic complications of Crohn's disease (CD) but their use as a primary therapy is more controversial, although this approach is frequently and successfully adopted in clinical practice. There is evidence that probiotic therapy may be effective in the prevention and treatment of mild to moderate UC. In contrast, a lack of successful study data at present precludes the widespread use of probiotics in the treatment of CD. Both antibiotics and probiotics appear to play a beneficial role in the treatment and prevention of pouchitis and further trials are warranted to fully quantify their clinical efficacy.

Animals↗

Do probiotics have a therapeutic role in gastroenterology?

Several hundred species of bacteria inhabit the gut, and affect its cell biology, morphology and homeostasis. Many bacteria are however potential pathogens, especially if the integrity of the epithelial barrier is physically or functionally breached. Conversely, the interaction between host and commensal microbes can confer important health benefits. This has led to commercial and public interest in "probiotics", live microbes principally taken as food supplements. Might probiotics also be used in disease therapy? Experimental evidence that probiotics modulate gut physiology, particularly barrier integrity and immunological function, underpins exciting new gastroenterological research. We discuss below the scientific basis for probiotic effects and present a critical perspective for their use in relation to gastrointestinal disease.

Gastrointestinal Diseases↗

Effects of probiotic bacteria on gastrointestinal motility in guinea-pig isolated tissue.

AIM: To evaluate the intestinal motility changes evoked by 8 bacterial strains belonging to Bifidobacterium, Lactobacillus and Streptococcus genera within the probiotic preparation VSL#3. METHODS: Ileum and proximal colon segments isolated from guinea-pigs were used as a study model. Entire cells and cell fractions (cell debris, cell wall fraction, cytoplasmatic fraction, proteinaceous and non-proteinaceous cytoplasmatic components) of VSL#3 strains and, as controls, Escherichia coli, Salmonella aboni and Bacillus licheniformis were tested in this in vitro model. RESULTS: Among the bacterial cell fractions tested, only the cytoplasmatic fraction modified intestinal motility. Lactobacillus strains stimulated the contraction of ileum segment, whereas all probiotic strains tested induced proximal colon relaxation response. The non-proteinaceous cytoplasmatic components were responsible for the colon relaxation. CONCLUSION: The results obtained in this study suggest that the proximal colon relaxation activity showed by the probiotic bacteria could be one of the possible mechanisms of action by which probiotics exert their positive effects in regulating intestinal motility.

Animals↗

Lysozyme-modified probiotic components protect rats against polymicrobial sepsis: role of macrophages and cathelicidin-related innate immunity.

Severe sepsis is associated with dysfunction of the macrophage/monocyte, an important cellular effector of the innate immune system. Previous investigations suggested that probiotic components effectively enhance effector cell functions of the immune system in vivo. In this study, we produced bacteria-free, lysozyme-modified probiotic components (LzMPC) by treating the probiotic bacteria, Lactobacillus sp., with lysozyme. We showed that oral delivery of LzMPC effectively protected rats against lethality from polymicrobial sepsis induced by cecal ligation and puncture. We found that orally administrated LzMPC was engulfed by cells such as macrophages in the liver after crossing the intestinal barrier. Moreover, LzMPC-induced protection was associated with an increase in bacterial clearance in the liver. In vitro, LzMPC up-regulated the expression of cathelicidin-related antimicrobial peptide (CRAMP) in macrophages and enhanced bactericidal activity of these cells. Furthermore, we demonstrated that surgical stress or cecal ligation and puncture caused a decrease in CRAMP expression in the liver, whereas enteral administration of LzMPC restored CRAMP gene expression in these animals. Using a neutralizing Ab, we showed that protection against sepsis by LzMPC treatment required endogenous CRAMP. In addition, macrophages from LzMPC-treated rats had an enhanced capacity of cytokine production in response to LPS or LzMPC stimulation. Together, our data suggest that the protective effect of LzMPC in sepsis is related to an enhanced cathelicidin-related innate immunity in macrophages. Therefore, LzMPC, a novel probiotic product, is a potent immunomodulator for macrophages and may be beneficial for the treatment of sepsis.

Animals↗

Probiotics, prebiotics and synbiotics: a role in chemoprevention for colorectal cancer?

Colorectal cancer (CRC) is the third most common form of cancer. Current treatments including chemotherapy, radiotherapy and surgery are all associated with a high risk of complications and are not always successful, highlighting the need to develop new treatment strategies. The ingestion of probiotics, prebiotics or combinations of both (synbiotics) represents a novel new therapeutic option. Probiotics and prebiotics act to alter the intestinal microflora by increasing concentrations of beneficial bacteria such as lactobacillus and bifidobacteria, and reducing the levels of pathogenic micro-organisms. This strategy has the potential to inhibit the development and progression of neoplasia via mechanisms including; decreased intestinal inflammation, enhanced immune function and anti-tumorigenic activity, binding to potential food carcinogens including toxins found in meat products, and a reduction in bacterial enzymes which hydrolyse precarcinogenic compounds, such as beta-glucuronidase. There is substantial experimental evidence to suggest that probiotics and prebiotics may be beneficial in the prevention and treatment of colon cancer, however to date there have been few conclusive human trials. Probiotics and prebiotics have the potential to impact significantly on the development, progression and treatment of colorectal cancer and may have a valuable role in cancer prevention.

Anticarcinogenic Agents↗

Evaluation of avian-specific probiotic and Salmonella enteritidis-, Salmonella typhimurium-, and Salmonella heidelberg-specific antibodies on cecal colonization and organ invasion of Salmonella enteritidis in broilers.

Salmonella Enteritidis colonizes the intestinal tract of poultry and causes foodborne illness in humans. Reduction of Salmonella Enteritidis colonization in the intestinal tract of poultry reduces potential carcass contamination during slaughter. The purpose of this study was to determine the effect of an avian-specific probiotic combined with Salmonella Enteritidis-, Salmonella Typhimurium-, and Salmonella Heidelberg-specific antibodies on the cecal colonization and organ invasion of Salmonella Enteritidis in broiler as well as on body weights. The treatment group was defined as chicks spray-vaccinated with Avian Pac Plus at the hatchery and given Avian Pac Plus for the first 3 days after placement. An intermediate treatment was given at 10 and 14 days, 2 days prior to vaccination and 2 days postvaccination. All birds were vaccinated with Newcastle disease vaccine, La Sota virus (one drop/eye) at 12 days of age. A final treatment was given 3 days preslaughter. The control group was defined as chicks not given Avian Pac Plus at any time. Six hours after oral administration of the probiotic suspension (treatment group) or water (control group) at placement, the chicks were challenged with Salmonella Enteritidis. All chickens were orally inoculated with 0.25 ml of Salmonella Enteritidis that contained 4 x 10(7) CFU/1.0 ml. Cecal colonization and organ invasion were evaluated for Salmonella Enteritidis on days 0, 1, 3, 7, 10, 17, 24, 31, 38, and 41. The probiotic-treated group had a significantly lower concentration of Salmonella Enteritidis cecal colonization at days 3, 7, 10, 17, 24, 31, 38, and 41 when compared to the nontreated, control group (P < 0.05). Similarly, there was a significant difference (P < 0.05) in the isolation of Salmonella Enteritidis from the internal organs (liver and spleen) when probiotic-treated and nonprobiotic-treated groups were compared. There was no significant difference (P > 0.05) in the mean body weight between the two experimental groups at each collection period. These results indicated that a combination of Lactobacillus acidophilus, Streptococcus faecium, and Salmonella Enteritidis, Salmonella Typhimurium, and Salmonella Heidelberg-Specific antibodies have a beneficial effect in reducing the colonization of Salmonella Enteritidis in market-aged broilers.

Animals↗

A washed-curd goat's cheese as a vehicle for delivery of a potential probiotic bacterium: Lactobacillus delbrueckii subsp. lactis UO 004.

This study characterizes the probiotic properties of Lactobacillus delbrueckii subsp. lactis UO 004 and examines its suitability for making cheese. This strain was isolated from infant feces and shows interesting features, such as acid and bile tolerance, adherence to intestinal epithelial cells, and inhibition of the growth of certain enteropathogens, that support its potential use as a probiotic strain. In this regard, the suitability of a washed-curd cheese (Vidiago type) made with goat's milk as a delivery system for this probiotic strain was assessed. Lactobacillus delbrueckii subsp. lactis UO 004 was incorporated into a starter culture (IPLA 001). Changes in the overall composition of control and experimental cheeses were determined during ripening through bacteriological, chemical, high-performance liquid chromatography, and gas chromatography analyses. Slight changes in the gross composition and appreciable differences in the flavor compounds profile were observed between control and experimental cheeses. This strain was capable of surviving at high cell numbers (10(8) to 10(9) CFU/g) in cheeses after 28 days of ripening without adversely affecting sensory criteria or appearance of the cheese, thus satisfying the criteria for a probiotic food product.

Animals↗

Assessment of the effects of Nurmi-type cultures and a defined probiotic preparation on a Salmonella typhimurium 29E challenge in vivo.

The effects of treatment with an undefined commercial Nurmi-type culture (NTC), cultured cecal contents, and a dual-strain probiotic, containing Enterococcus faecalis and Pediococcus pentosaceus, on Salmonella Typhimurium colonization were evaluated in a specific-pathogen-free bird model. Two sets of trials were performed, and each study was arranged as a randomized complete block design with three treatments. Treatments consisted of (i) control, (ii) commercial NTC, and (iii) cultured cecal contents in the first set of trials and (i) control, (ii) defined probiotic, and (iii) cultured cecal contents in the second set. On day 1, birds were administered 1.2 x 10(7) CFU of the appropriate treatment by oral gavage. On day 3, all birds were challenged with 1 x 10(6) CFU of Salmonella Typhimurium 29E (nalidixic acid resistant). Chicks were asphyxiated with argon gas on day 10, and ceca were aseptically removed. Salmonella Typhimurium counts (CFU per milliliter of cecal contents) were determined on brilliant green agar containing 30 mg of nalidixic acid per liter, and CFU counts were log transformed prior to analysis. Cecal pH and volatile fatty acid concentrations were also determined. Data were analyzed by one-way analysis of variance, and means were compared by Tukey's pairwise analysis. Commercial NTC and cultured cecal contents treatments resulted in a significant decrease (P < or = 0.05) in Salmonella Typhimurium 29E colonization, with the NTC offering a higher level of protection. In the second set of trials, the defined probiotic tended to reduce colonization by Salmonella Typhimurium (P = 0.07), while chicks treated with cultured cecal contents displayed a significant decrease (P = 0.03) when compared to the negative control. No significant change was observed in cecal pH or in acetate and propionate concentrations; however, a significant increase in butyrate concentrations in both the cultured cecal contents and defined probiotic treatment groups was observed when compared to the control birds. These observations suggest that defined cultures are less effective Salmonella control agents than are preparations generated from the complete cecal microflora.

Administration, Oral↗

[Combined antibacterial and probiotic therapy of Helicobacter-associated diseases in children].

On the basis of microbiological monitoring and clinico-morphological criteria, the use of probiotics containing lacto- and bifidobacteria, simultaneously with "triple" antibacterial therapy (antibiotics of choice, metronidazole and bismuth salts), has been found to produce curative effect in the treatment of Helicobacter-associated gastroduodenal pathology in children. The influence of "triple" antibacterial therapy on the microecology of the gastrointestinal tract and the state of the immunobiological status of patients was studied. The combined scheme for correcting damaged microbiocenosis with probiotics in accordance with the concept of microecological adequacy is proposed. The scheme includes modern antibacterial therapy in combination with the probiotic preparations of lactobacteria. The prescription of bifidobacteria-containing probiotics is recommended at early stages from the beginning of etiotropic therapy. The effectiveness of the therapy may be dynamically evaluated, starting from week 5 after the end of the complex therapy, in EIA with the use of specific anti-Helicobacter antibodies.

Anti-Bacterial Agents↗

Probiotics: established effects and open questions.

Human intestinal microflora is a complex ecosystem with hundreds of bacterial species. Its metabolic functions and interactions with the host probably affect the human health and well being, but these effects are extremely difficult to study. However, for about 100 years, the idea of modifying the composition of colonic flora by consuming viable bacteria in order to improve the quality of life and to prevent and treat intestinal disorders has had some popularity. Solid clinical data have usually been lacking to support the health claims associated with these so-called probiotics. The situation, however, is rapidly changing. Competent clinical studies are accumulating, showing that specific probiotic microbes, mainly lactic acid bacteria and bifidobacteria, can alleviate or prevent diverse intestinal disorders and reduce the risk of some intestinal diseases. Some indication of the mechanisms of action can also be deduced from the data available, while rapidly developing molecular biological methods offer new tools to verify the survival of the probiotics in the gut and the subsequent adhesion to mucosae. While development of foods containing probiotic bacteria has a great potential for the food industry and can be expected to positively affect the general health of the population, safety considerations have to be taken into account while introducing new species or strains without a previous history of safe food use.

Bifidobacterium↗