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Comparison of four methods to enumerate probiotic bifidobacteria in a fermented food product.

Four methods of enumeration were compared by monitoring levels of probiotic bifidobacteria in fermented oat drink during storage. Strains of Bifidobacterium longum and B. lactis were quantified by plate counts, fluorescent in situ hybridization (FISH), quantitative real-time PCR and commercial LIVE/DEAD BacLight bacterial viability kit, and the methods were further developed to suit the enumeration of bifidobacteria in fermented foods. Plate counts of both B. lactis and B. longum were lower than the PCR and FISH counts. The LIVE/DEAD counts of B. lactis were comparable to PCR and FISH counts. The plate counts of B. lactis were slightly but significantly lower than LIVE/DEAD counts, suggesting that the cells that were not able to grow on plates may have become dormant. The plate counts of B. longum were several log units lower than LIVE/DEAD counts, suggesting that a remarkable part of the cells were dormant. Real-time PCR and FISH were shown to suit the quantification of the total amount of probiotic bifidobacteria in foods. Plate counts and LIVE/DEAD counts provided conflicting information on viability especially in the case of B. longum. We conclude that the choice of enumeration method for probiotic bacteria may have significant effect on the results of the analysis. The strain-specific properties and the objects of the analysis should be taken into account when enumeration methods for different probiotic strains are chosen.

Bifidobacterium↗

Antibiotic susceptibility profiles of new probiotic Lactobacillus and Bifidobacterium strains.

The antimicrobial susceptibilities and presence of plasmids in four new probiotic lactic acid bacteria (LAB) strains, Lactobacillus rhamnosus HN001 (DR20) HN067, Lactobacillus acidophilus HN017 and Bifidobacterium lactis HN019 (DR10), were determined. Resistance to 18 commonly used antibiotics was assessed by disk diffusion. The three Lactobacillus strains had similar antibiotic susceptibility profiles to those of Lactobacillus plantarum strain HN045 and two commercial probiotic Lactobacillus strains, GG and LA-1. The B. lactis strain HN019 had a similar profile to three commercial probiotic B. lactis strains (Bb12, HN049 and HN098). All 10 strains were sensitive to the Gram-positive spectrum antibiotics erythromycin and novobiocin, the broad-spectrum antibiotics rifampicin, spectinomycin, tetracycline and chloramphenicol and the beta-lactam antibiotics penicillin, ampicillin and cephalothin. By contrast, most strains were resistant to the Gram-negative spectrum antibiotics fusidic acid, nalidixic acid and polymyxin B and the aminoglycosides neomycin, gentamicin, kanamycin and streptomycin. All three L. rhamnosus strains (HN001, HN067 and GG) were resistant to vancomycin and several strains were also resistant to cloxacillin. Of the four new probiotic strains, only L. rhamnosus HN001 contained plasmids; however, a plasmid-free derivative of HN001 had the same antibiotic susceptibility profile as the parent strain.

Anti-Bacterial Agents↗

Immunostimulatory probiotic Lactobacillus rhamnosus HN001 and Bifidobacterium lactis HN019 do not induce pathological inflammation in mouse model of experimental autoimmune thyroiditis.

The possibility that intestinal microflora contribute to the pathogenesis of autoimmune diseases has raised issues regarding the safety of probiotic organisms, especially those with immunostimulating properties, in individuals with such immune dysfunctions. In this study, the effect of consumption of probiotic lactic acid bacteria (LAB) strains Lactobacillus rhamnosus HN001(HN001) and Bifidobacterium lactis HN019 (HN019) on the induction and progression of experimental autoimmune thyroiditis (EAT) was investigated in CBA/CaH (H-2k) mice. HN001 or HN019 in skim milk were fed to mice daily (1-1.5 x 10(8) cfu/mouse/day) for 5 to 9 weeks. A mild form of EAT was induced by subcutaneous injection of mouse thyroglobulin (MTg) with either Freund's adjuvant (complete and incomplete, CFA and IFA) or lipopolysaccharide (LPS). The proliferative responses of spleen lymphocyte to MTg stimulation in vitro and the presence (and degree) of mononuclear cell infiltration in thyroid gland tissues were examined to assess the development and severity of EAT. The levels of serum anti-MTg antibodies (IgG1 and IgG2a) and spleen weight index were determined to detect the presence of autoimmune responses of mice receiving MTg. Results showed that 8 weeks after immunization, 16.67-50% of the mice developed mild EAT with lymphocyte infiltration in the thyroid glands. Probiotic feeding did not induce full-blown EAT. There were no differences in spleen weight index or the proliferative spleenocytes in response to PMA between mice that received MTg alone and mice that received MTg and probiotic LAB strains.

Animals↗

A probiotic strain of Lactobacillus plantarum stimulates lymphocyte responses in immunologically intact and immunocompromised mice.

Experimental evidences showing the immunomodulatory effects of probiotic microorganisms have been provided by studies on immunologically intact animals. Here we compared the immunomodulation capacity of a probiotic strain of Lactobacillus plantarum on intact and cyclophosphamide-treated BALB/c mice. Although this strain fulfilled the in vitro criteria for the selection of potentially probiotic bacteria (resistance to low pH and bile, adhesion to epithelial cells and antimicrobial activity), it was unable to establish a persistent colonization in the gastrointestinal tract after intragastric gavage. The administration of L. plantarum did not modify the cyclophosphamide-induced leukopenia, but partially restored the proliferation of spleen cells from cyclophosphamide-treated mice in response to lipopolysaccharide. Our findings show that probiotic bacteria may exert immunomodulatory effects despite a limited colonization ability and may improve the immune function damaged by immunosuppressive agents.

Administration, Oral↗

Lactobacillus fermentum BR11, a potential new probiotic, alleviates symptoms of colitis induced by dextran sulfate sodium (DSS) in rats.

Current treatments for inflammatory bowel disease (IBD) are relatively ineffective. Recently, probiotics have emerged as a potential treatment modality for numerous gastrointestinal disorders, including IBD. Few probiotics, however, have undergone appropriate preclinical screening in vivo. The current study compared the effects of four candidate probiotics on development of dextran sulfate sodium (DSS)-induced colitis in rats. Sprague Dawley rats were gavaged 1 mL of the potential probiotic (1 x 10(10) CFU/mL), or vehicle, twice daily for 14 days. Strains tested were Lactobacillus rhamnosus GG (LGG), Streptococcus thermophilus TH-4 (TH-4), Bifidobacterium lactis Bb12 (Bb12) and Lactobacillus fermentum BR11 (BR11). Colitis was induced from day 7 to 14 via administration of 2% DSS in drinking water. Disease activity index (DAI) was monitored daily until rats were killed at day 14. DAI decreased in DSS+Bb12 and DSS+BR11 compared to DSS+Vehicle. Colon length increased in DSS+BR11 (10%) and DSS+LGG (10%) compared to DSS+Vehicle. DSS+Bb12 and DSS+BR11 prevented the distal colon crypt hyperplasia evident in DSS+Vehicle, DSS+LGG and DSS+TH-4. BR11 was most effective at reducing colitic symptoms. Bb12 had minimal effects, whilst TH-4 did not prevent DSS-colitis and LGG actually exacerbated some indicators of colitis. Further studies into the potential benefits of L. fermentum BR11 are indicated.

Animals↗

Effect of probiotics on preventing disruption of the intestinal microflora following antibiotic therapy: a double-blind, placebo-controlled pilot study.

In this pilot-scale, double-blind, placebo-controlled trial, 30 patients with Helicobacter pylori infection were randomised into three groups prior to their 7 days eradication therapy, to study the effects of probiotic supplement comprising Lactobacillus acidophilus and Bifidobacterium bifidum on the intestinal microflora in response to antibiotic therapy. Group I received the placebo product from day 1 to day 15, Group II received placebo from day 1 to day 7 and probiotics from day 8 to day 15 and Group III received probiotics from day 1 to day 15. Patients provided stool samples for analysis on days 1, 7, 12, 17 and 27. For patients in Groups I and II, significant increases in the facultative anaerobe component of the microflora occurred between days 1 and 7. In Group I, the numbers remained elevated to day 27 but in Group II, the numbers decreased significantly between days 7 and 27 back to the starting levels. In Group III, the facultative anaerobe population remained stable throughout. The total anaerobe numbers increased significantly at day 27 than at day 1 for Group I, were unchanged throughout for Group II and decreased significantly for the patients in Group III between days 1 and 7 before reverting to the starting levels by day 27. From these results, it can be seen that probiotic supplementation modulates the response of the intestinal microflora to the effects of antibiotic therapy.

Aged↗

Selective probiotic bacteria induce IL-10-producing regulatory T cells in vitro by modulating dendritic cell function through dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin.

BACKGROUND: Lactobacilli are probiotic bacteria that are frequently tested in the management of allergic diseases or gastroenteritis. It is hypothesized that these probiotics have immunoregulatory properties and promote mucosal tolerance, which is in part mediated by regulatory T cells (Treg cells). On the basis of pathogenic or tissue-specific priming, dendritic cells (DC) acquire different T cell-instructive signals and drive the differentiation of naive T H cells into either T H 1, T H 2, or regulatory effector T cells. OBJECTIVE: We studied in what way different species of lactobacilli prime human DCs for their ability to drive Treg cells. METHODS: Human monocyte-derived DCs were cultured in vitro with lactobacilli of different species. RESULTS: Two different species of lactobacilli, Lactobacillus reuteri and Lactobacillus casei , but not Lactobacillus plantarum, prime monocyte-derived DCs to drive the development of Treg cells. These Treg cells produced increased levels of IL-10 and were capable of inhibiting the proliferation of bystander T cells in an IL-10-dependent fashion. Strikingly, both L reuteri and L casei , but not L plantarum , bind the C-type lectin DC-specific intercellular adhesion molecule 3-grabbing non-integrin (DC-SIGN). Blocking antibodies to DC-SIGN inhibited the induction of the Treg cells by these probiotic bacteria, stressing that ligation of DC-SIGN can actively prime DCs to induce Treg cells. CONCLUSIONS: The targeting of DC-SIGN by certain probiotic bacteria might explain their beneficial effect in the treatment of a number of inflammatory diseases, including atopic dermatitis and Crohn's disease.

Antigens, CD↗

Modulation of carbohydrate metabolism and peptide hormones by soybean isoflavones and probiotics in obesity and diabetes.

Soybean and its isoflavones have been shown to have beneficial effects on carbohydrate and lipid metabolism and on renal function. Probiotics may potentiate the beneficial effects of isoflavones by converting the inactive isoflavone glycoside to aglycones, which are biologically active, thereby producing a synergistic effect. We therefore studied the effects of soybean isoflavones in the presence and absence of probiotics on glucose and triglyceride metabolism and the peptide hormones involved in their metabolism. Lean and obese SHR/N-cp rats were fed AIN-93 diets containing 0.1% soybean isoflavone mixture, 0.1% probiotics mixture or both. Plasma was analyzed for glucose, triglycerides, parameters of renal function and peptide hormones -- insulin, leptin, glucagon and ACTH -- that are involved in glucose and lipid metabolism. Isoflavones given alone lowered plasma glucose in both phenotypes while triglyceride was decreased only in lean animals. Isoflavones also lowered aspartate amino transferase and alanine amino transferase in both phenotypes. Isoflavones had significant effect on plasma insulin, leptin and glucagon in lean rats but not in obese rats. Thus, our data show that in lean animals, isoflavones have hypoglycemic and hypolipidemic effect, and the effect is mediated by changes in peptide hormones. When lipid levels are very high as in obese rats, isoflavones fail to lower plasma triglyceride levels. Probiotics do not appear to enhance the effect of isoflavones.

Adrenocorticotropic Hormone↗

Effect of probiotics on gastrointestinal symptoms and small intestinal permeability in children with atopic dermatitis.

OBJECTIVE: To determine whether probiotic lactobacilli may alleviate small intestinal inflammation and strengthen the intestinal barrier function in children with atopic dermatitis. STUDY DESIGN: In a double-blinded, placebo-controlled, cross-over study, probiotic lactobacilli (Lactobacillus rhamnosus 19070-2 and L reuteri DSM 12246) were administered for 6 weeks to 41 children with moderate and severe atopic dermatitis. Gastrointestinal symptoms were registered before and during treatment and small intestinal permeability was measured by the lactulose-mannitol test. RESULTS: During Lactobacillus supplementation, there was a significant decrease in the frequency of gastrointestinal symptoms (39% during the placebo period versus 10% during active treatment, P=.002). There was a positive association between the lactulose to mannitol ratio and the severity of the eczema (r=0.61, P=.02 after placebo and r=0.53, P=.05 after active treatment). After probiotic treatment, the lactulose to mannitol ratio was lower (0.073) than after placebo (0.110, P=.001). CONCLUSIONS: Impairment of the intestinal mucosal barrier appears to be involved in the pathogenesis of atopic dermatitis. The study suggests that probiotic supplementation may stabilize the intestinal barrier function and decrease gastrointestinal symptoms in children with atopic dermatitis.

Adolescent↗

Oral probiotics prevent necrotizing enterocolitis in very low birth weight neonates.

OBJECTIVE: To test the hypothesis that normalizing the intestinal flora by administration of prophylactic probiotics would provide a natural defense, thereby reducing both the incidence and severity of necrotizing enterocolitis (NEC) in preterm neonates. STUDY DESIGN: Neonates < or =1500 g birth weight were randomized to either receive a daily feeding supplementation with a probiotic mixture (Bifidobacteria infantis, Streptococcus thermophilus, and Bifidobacteria bifidus; Solgar, Israel) of 10(9) colony forming units (CFU)/day or to not receive feed supplements. NEC was graded according to Bell's criteria. RESULTS: For 72 study and 73 control infants, respectively, birth weight (1152 +/- 262 g vs 1111 +/- 278 g), gestational age (30 +/- 3 weeks vs 29 +/- 4 weeks), and time to reach full feeds (14.6 +/- 8.7 days vs 17.5 +/- 13.6 days) were not different. The incidence of NEC was reduced in the study group (4% vs 16.4%; P=.03). NEC was less severe in the probiotic-supplemented infants (Bell's criteria 2.3 +/- 0.5 vs 1.3 +/- 0.5; P=.005). Three of 15 babies who developed NEC died, and all NEC-related deaths occurred in control infants. CONCLUSION: Probiotic supplementation reduced both the incidence and severity of NEC in our premature neonatal population.

Administration, Oral↗

Effect of a probiotic food as an adjuvant to triple therapy for eradication of Helicobacter pylori infection in children.

OBJECTIVE: Current recommendations for treatment of Helicobacter pylori infection include a proton pump inhibitor in combination with two antibiotics. We evaluated the potential activity of a probiotic food as an adjuvant to antibiotic triple therapy for eradication of H. pylori infection in children from Buenos Aires, Argentina. METHODS: Sixty-five children who tested positive for H. pylori, as diagnosed by (13)C-urea breath test and endoscopy, were included in this study. Patients were randomized to receive 1-wk triple therapy plus probiotic food (treated group) or milk placebo (control) that was administered for 3 mo. Probiotic food consisted of 250 mL of a commercial yogurt containing Bifidobacterium animalis and Lactobacillus casei (10(7) colony-forming units/mL). Post-treatment urea breath test controls were performed 1 and 3 mo after the end of triple therapy. RESULTS: We found no significant differences in H. pylori eradication rates (ERs) at 1 and 3 mo between the treated group (ER = 45.5% and 42.4%) and the control group (ER = 37.5% and 40.6%). Relative risks between groups were 0.87 (95% confidence interval 0.58-1.32, P = 0.345) in the first month and 0.97 (95% confidence interval 0.64-1.46, P = 0.542) in the third month. CONCLUSIONS: We could not demonstrate an adjuvant effect of the studied probiotic food to triple therapy in the eradication of H. pylori infection in children in Buenos Aires, Argentina. However, we found lower ERs than those reported for the same therapeutic scheme in developed countries, indicating that bacterial resistance and alternative therapeutic strategies should be studied.

Adolescent↗

Safe and efficacious probiotics: what are they?

Each year, >20 billion doses of probiotics are used by healthy people and by those diagnosed with a range of medical conditions. Compared to many pharmaceutical agents, probiotics are well tolerated and extremely safe, and serious adverse effects rarely occur. Nevertheless, as many new researchers enter the field and companies launch "probiotic" products, it is essential that standards are set for naming a product "probiotic" to show that it meets an acceptable level of safety and efficacy, and to understand the strengths and limitations of its activity. In this Opinion article, recommendations are made based upon the current understanding of scientific, clinical and regulatory issues, with a special focus on safety.

Animals↗

Effect of orally administered probiotic E. coli strain Nissle 1917 on intestinal mucosal immune cells of healthy young pigs.

Several beneficial effects of probiotics have been described in studies using rodent disease models and in human patients; however, the underlying mechanisms remained mostly unclear. Only a few studies focused on the effects of probiotics on the intestinal mucosal immune system. Here, we studied the effect of the probiotic strain E. coli Nissle 1917 (EcN) administered orally to young pigs at two concentrations (10(9) and 10(11)CFU/d for 21 days) on the gut-associated lymphatic tissue. This probiotic strain was shown recently to reduce recurrence of inflammation in ulcerative colitis patients. We quantified the number and distribution of intestinal immune cells (granulocytes, mast cells, CD4+, CD8+, CD25+, IgA+ lymphocytes) and the mucosal mRNA expression of cytokines (IFN-gamma, TNF-alpha, TGF-beta, IL-10) and antimicrobial peptides (PR-39, NK-lysin, prepro-defensin-beta 1, protegrins). The number and distribution of cells were highly different between small intestinal and colon segments in all groups, but were not influenced by EcN, except high dose EcN fed pigs (10(11) CFU/d) showing an increase in mucosal CD8+ cells in the ascending colon. The mRNA analysis revealed no changes associated with EcN feeding. In conclusion, according to our analyses EcN has only minor effects on the distribution of mucosal immune cells in the gut of healthy individuals. The well-established preventive effects of EcN might therefore be relate to other mechanisms than simple modulation of immune cell distribution.

Animals↗

Does probiotics administration decrease serum endotoxin levels in infants?

PURPOSE: The aim of this study was to examine whether administration of probiotics to infants can change the ratio of intestinal flora and thereby decrease serum endotoxin produced by potentially pathogenic microorganisms. METHODS: Nine infants including five with of biliary atresia, two with omphalocele, one each with Hirschsprung's disease and imperforate anus were studied. All patients were stable, and no antibiotics were given during this study. A probiotic mixture consisting of Streptococcus faecalis, Clostridium butyricum and Bacillus mesentericus was administered orally to each infant at 2 g/day for 2 weeks. Fecal aerobic and anaerobic bacterial cultures, serum endotoxin level, and other biochemical parameters were examined. RESULTS: In fecal cultures, anaerobic bacteria including Bifidobacterium increased significantly whereas Escherichia coli, Streptococcus, and Klebsiella tended to decrease. The ratio of anaerobic to aerobic bacteria increased five times as a result of administration of probiotics, and serum endotoxin levels decreased. CONCLUSIONS: Probiotics affect intestinal bacterial flora by increasing anaerobic bacteria and decreasing the population of potentially pathogenic microorganisms. A decrease in luminal endotoxin may result in less endotoxin translocation or bacterial translocation.

Administration, Oral↗

Assessment of adhesion properties of novel probiotic strains to human intestinal mucus.

Potential new probiotic strains Lactobacillus brevis PELI, L. reuteri ING1, L. rhamnosus VTT E-800 and L. rhamnosus LC-705 were assessed for their adhesion properties using the human intestinal mucus model. The effect on the adhesion of exposure to acid and pepsin and to milk were tested to simulate gastric and food processing conditions, and the effect of different growth media on adhesion was tested. The properties of the four strains were compared to the well-investigated probiotic L. rhamnosus strain GG. Three of the tested strains showed significant adhesion properties in the mucus model, while L. brevis PELI had intermediate adhesion and L. rhamnosus LC-705 adhered poorly. Pretreatment with different milks decreased the adhesion and low pH and pepsin treatment reduced the adhesion of all tested strains except L. rhamnosus LC-705. No competitive exclusion of pathogenic Salmonella typhimurium or Escherichia coli SfaII was observed. The results indicate that major differences exist between tested proposed probiotic strains. The growth media and the food matrix significantly affect the adhesive ability of the tested strains. This has previously not been taken into account when selecting novel probiotic strains.

Acids↗

Growth and metabolism of selected strains of probiotic bacteria in milk.

Growth and metabolism of five probiotic strains with well-documented health effects were studied in ultra-high temperature (UHT) treated milk, supplemented with 0.5% (w/v) tryptone or 0.75% (w/v) fructose. The probiotic strains were Lactobacillus acidophilus La5, Lb. acidophilus 1748, Lactobacillus rhamnosus GG, Lactobacillus reuteri SD 2112 and Bifidobacterium animalis BB12. Fermentation was followed for 72 h at 37 degrees C and the samples were analysed for pH, log cfu ml(-1), volatile compounds, organic acids and carbon dioxide. The strains reduced pH from 6.7 to between 3.9 and 4.4 after 24 h of incubation. All strains attained viable cell counts above 8.7-9.18 log cfu ml(-1) after 6-16 h of incubation. The two Lb. acidophilus strains showed a stable level of viable cells during 12-72 h of incubation but the three other strains showed a reduction of 0.4-1.1 log cfu ml(-1) from 24 to 72 h of incubation. However, all strains showed cell levels between 7.8 and 8.7 log cfu ml(-1) after 72 h of incubation. After 48 h of incubation, the amount of lactic acid produced varied according to strain from 6949 to 14,000 mg kg(-1) and acetic acid produced varied from 0 to 6901 mg kg(-1). Three of the strains metabolised citrate but only low amounts of diacetyl and acetoin were detected within strains, 0.2-0.8 and 6.5-10 mg kg(-1), respectively. Carbon dioxide produced varied from 221 to 3942 mg kg(-1) and was connected to the citrate-fermenting ability of the strain used and their carbohydrate fermentation pathway. Three of the strains produced detectable levels of acetaldehyde and the concentration varied from 9.4 to 12.6 mg kg(-1) after 24 h of incubation. All five probiotic strains showed very different profiles of metabolites during fermentation; however, the two Lb. acidophilus strains were the most alike. Our findings show the importance of controlling the fermentation time since the probiotic strains produced different amounts of metabolic products according to fermentation time.

Animals↗

Demonstration of safety of probiotics -- a review.

Probiotics are commonly defined as viable microorganisms (bacteria or yeasts) that exhibit a beneficial effect on the health of the host when they are ingested. They are used in foods, especially in fermented dairy products, but also in pharmaceutical preparations. The development of new probiotic strains aims at more active beneficial organisms. In the case of novel microorganisms and modified organisms the question of their safety and the risk to benefit ratio have to be assessed. Lactic acid bacteria (LAB) in foods have a long history of safe use. Members of the genera Lactococcus and Lactobacillus are most commonly given generally-recognised-as-safe (GRAS) status whilst members of the genera Streptococcus and Enterococcus and some other genera of LAB contain some opportunistic pathogens. Lactic acid bacteria are intrinsically resistant to many antibiotics. In many cases resistances are not, however, transmissible, and the species are also sensitive to many clinically used antibiotics even in the case of a lactic acid bacteria- associated opportunistic infection. Therefore no particular safety concern is associated with intrinsic type of resistance. Plasmid-associated antibiotic resistance, which occasionally occurs, is another matter because of the possibility of the resistance spreading to other, more harmful species and genera. The transmissible enterococcal resistance against glycopeptide antibiotics (vancomycin and teicoplanin) is particularly noteworthy, as vancomycin is one of the last effective antibiotics left in the treatment of certain multidrug-resistant pathogens. New species and more specific strains of probiotic bacteria are constantly identified. Prior to incorporating new strains into products their efficacy should be carefully assessed, and a case by case evaluation as to whether they share the safety status of traditional food-grade organisms should be made. The current documentation of adverse effects in the literature is reviewed. Future recommendations for the safety of already existing and new probiotics will be given.

Animals↗

The effect of probiotic strains on the microbiota of the Simulator of the Human Intestinal Microbial Ecosystem (SHIME).

The aim of the present work was to study five potential probiotic strains (Lactobacillus plantarum, two strains of L. paracasei subsp. paracasei, L. rhamnosus and Bifidobacterium sp.) comparatively in the Simulator of the Human Intestinal Microbial Ecosystem (SHIME) in vitro model, and to evaluate this model as a tool in the screening and selection of probiotic bacteria. The impact of the strains on the composition of microbiota and its metabolic activities (production of lactic acid and short-chain fatty acids) was studied. Changes in composition of the microbiota become apparent as a result of probiotic treatment. A marked, but temporary, increase was noted in the number of lactic acid bacteria and bifidobacteria. The profiles of D(-) and L(+) isomers of lactic acid detected in the SHIME after addition of probiotic strains corresponded well to those that are produced in pure culture conditions. The numbers of enterobacteriaceae decreased markedly and those of clostridia detectably during the intervention, while the enterococci tended to increase after the treatment. This pattern was similar in the reactors representing both the small and large intestine in the model. The changes in short-chain fatty acids were small, and no definite trend was observed.

Bifidobacterium↗