At Ethical, we’re proud to be honest and transparent with our products and what’s in them. And we’re happy to share our Probiotic study with you.
Probiotic Capsule – White Paper v.1
- Supplement Facts/ Product Formulation
| Supplement Facts | ||
| Serving Size 2 veggie capsules Servings Per Container 30 | ||
| Amount Per Serving | % Daily Value | |
| Bacillus subtilis DE111® | 5 billion CFU | * |
| Fructooligosaccharide (FOS) prebiotic fiber | 1 g | * |
| * Daily value not established. | ||
| Other ingredients: TBD |
DE111® is a registered trademark of Deerland Enzymes.®
- Directions
Take two capsules daily, preferably on an empty stomach.
- Claims
| Claim | Substantiation |
| Overall health claims | |
| Bacillus subtilis is a spore-forming probiotic that provides its own protection against destruction by stomach acids and survive transit through the stomach intact. † | Bacillus subtilis survival study 1DE111 gastrointestinal survival study 1 |
| Bacillus subtilis DE111 has been shown to survive stomach acid and bile salt conditions intact. † | DE111 gastrointestinal survival study 1 |
| This pre- and probiotic product is formulated to promote a healthy microflora in the gut and support overall health. † orThis probiotic dietary supplement helps support overall health. † | Bacillus subtilis DE111 study 1DE111 safety and efficacy study 1DE111 gastrointestinal health study 1DE111 gastrointestinal and immune study 1DE111 cardiovascular study 1 |
| Bacillus subtilis DE111 supports healthy digestion, a healthy immune system, and proper metabolism of the components of fat. † | Bacillus subtilis DE111 study 1DE111 safety and efficacy study 1DE111 gastrointestinal health study 1 |
| Digestion/gut claims | |
| Bacillus subtilis DE111 supports healthy gut functions. † | Bacillus subtilis DE111 study 1DE111 safety and efficacy study 1DE111 gastrointestinal health study 1 |
| Bacillus subtilis DE111 stimulates normal microflora in the gut. † | Bacillus subtilis DE111 study 1DE111 safety and efficacy study 1 |
| Bacillus subtilis DE111 supports a healthy balance between desirable and undesirable bacteria in the gut. † | Bacillus subtilis DE111 study 1DE111 safety and efficacy study 1 |
| Bacillus subtilis DE111 was clinically tested to help increase levels of healthy Bifidobacteria in the gut. † | Bacillus subtilis DE111 study 1DE111 safety and efficacy study 1 |
| Bacillus subtilis DE111 promotes digestive health. † | Bacillus subtilis DE111 study 1DE111 safety and efficacy study 1DE111 gastrointestinal health study 1 |
| Bacillus subtilis DE111 supports normal digestion. † | Bacillus subtilis DE111 study 1DE111 safety and efficacy study 1DE111 gastrointestinal health study 1 |
| Bacillus subtilis DE111 promotes a healthy GI tract. † | Bacillus subtilis DE111 study 1DE111 safety and efficacy study 1DE111 gastrointestinal health study 1 |
| Bacillus subtilis DE111 promotes digestive health. † or Bacillus subtilis DE111 supports normal digestive. † | DE111 gastrointestinal health study 1 |
| Bacillus subtilis DE111 supports regularity. † orBacillus subtilis DE111 supports normal bowel movements. † | DE111 gastrointestinal health study 1 |
| Bacillus subtilis DE111 helps protect against occasional constipation or diarrhea. † | DE111 gastrointestinal health study 1 |
| Cardiometabolic claims | |
| Bacillus subtilis DE111 was clinically tested to support healthy cholesterol levels already within normal ranges. † | Bacillus subtilis DE111 study 1 |
| Bacillus subtilis DE111 was clinically tested to support healthy triglyceride levels already within normal ranges. † | Bacillus subtilis DE111 study 1DE111 safety and efficacy study 1 |
| Bacillus subtilis DE111 supports heart health. † orBacillus subtilis DE111 promotes a healthy heart. † orBacillus subtilis DE111 supports cardiovascular health. † | DE111 cardiovascular study 1 |
| Bacillus subtilis DE111 supports healthy blood flow. † | DE111 cardiovascular study 1 |
| Bacillus subtilis DE111 was clinically tested to support healthy glucose levels already within normal ranges. † | Bacillus subtilis DE111 study 1DE111 safety and efficacy study 1 |
| Children claims | |
| Bacillus subtilis DE111 promotes healthy microbiome diversity in young children. † or Bacillus subtilis DE111 promotes a healthy microbiome in young children. † | DE111 children study 1 |
| Bacillus subtilis DE111 promotes intestinal health in young children. † | DE111 children study 1-2 |
| Bacillus subtilis DE111 promotes healthy immune function in young children. † | DE111 children study 2 |
| Bacillus subtilis DE111 promotes healthy digestion in young children. † | DE111 children study 2 |
| Bacillus subtilis DE111 supports microbiome diversity in children that promotes healthy digestion and immune functions. † | DE111 children study 1-2 |
| Bacillus subtilis DE111 supports normal bowel movements in young children. † orPromotes healthy bowel movements in young children. † | DE111 children study 2 |
| Bacillus subtilis DE111 promotes gastrointestinal health in young children. † | DE111 children study 1-2 |
| Immune claims | |
| Bacillus subtilis DE111 supports healthy immune health. † | Bacillus subtilis DE111 study 1 |
| Bacillus subtilis DE111 supports robust immune system function that naturally supports good health. † | DE111 gastrointestinal and immune study 1 |
| Bacillus subtilis DE111 promotes a healthy immune system and response that helps to support good health naturally. † | DE111 gastrointestinal and immune study 1 |
| Body composition claims | |
| Bacillus subtilis DE111 supports normal fat metabolism. † | |
| Bacillus subtilis DE111 helps optimize body composition with strength training. † | DE111 athlete study 2 |
| Bacillus subtilis DE111 supports improved results and performance with strength training.† | DE111 athlete study 2 |
| Bacillus subtilis DE111 supports exercise recovery. † | DE111 athlete study 2 |
| Bacillus subtilis DE111 was shown to help induce an inflammatory marker associated with exercise. † | DE111 athlete study 1 |
| As part of strength training program, Bacillus subtilis DE111 was shown in a clinical trial with athletes to support a leaner body and improved strength. † | DE111 athlete study 2 |
| FOS claims | |
| FOS helps support the growth of the desirable, probiotic bacteria in the intestines.† | FOS study 1-2, 3-6 |
†These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure, or prevent any disease.
- Substantiation Studies
Bacillus subtills survival study 1
As part of an ongoing study to determine the true habitat of Bacillus species, we report here the isolation and characterisation of Bacillus subtilis from the human gastrointestinal tract (GIT). Strains were obtained from ileum biopsies as well as from faecal samples and their biotypes defined. 16S rRNA analysis revealed that most isolates of B. subtilis were highly conserved, in contrast to RAPD-PCR fingerprinting that showed greater diversity with 23 distinct RAPD types. The majority of B. subtilis strains examined possessed features that could be advantageous to survival within the GIT. This included the ability to form biofilms, to sporulate anaerobically and secretion of antimicrobials. At least one isolate was shown to form spores that carried an exosporium, a loosely attached outer layer to the mature endospore, this being the first report of B. subtilis spores carrying an exosporium. This study reinforces a growing view that B. subtilis and probably other species have adapted to life within the GIT and should be considered gut commensals rather than solely soil microorganisms.
DE111 gastrointestinal survival study 1
A study was conducted to examine the pH survivability of Bacillus subtilis DE111. Gastric fluid simulation was as follows: The United States Pharmacopeia (USP 32: Dietary Supplements)/<2040> Disintegration and Dissolution of Dietary Supplements 1782 was performed in order to establish the survivability of DE111 under acidic conditions. Simulated gastric fluid was prepared, with and without pepsin, at a final pH of 1.2. DE111 (100 billion CFU/gram) was inoculated in both gastric fluid preparations and incubated at 37 ± 1°C for 1 hour. Cultures were serially diluted, inoculated on 3M plates, and incubated at 37 ± 1°C for 24 hours. As seen in the bar graph below, DE111 viability was maintained under USP 32/<2040>/1782 acidic conditions.
Acidic and bile salts conditions (24 hours) were as follows: The survivability of DE111 in acidic and bile salt conditions was assayed following the publication by Jiang, et al. The method for tolerance to acid and bile salt concentration was done with some minor modifications. Overnight cultures of DE111 (100 billion CFU/gram) in nutrient broth (24 h) were inoculated in nutrient broths that were adjusted to pH 4.5, 3.5, and 2.5 with HCl (1.0 M) and in non-acidified broth (pH 6.9) which served as a control. To test for bile salt survivability, cultures were inoculated into nutrient broths supplemented with 0.15, 0.30, and 0.45% (wt/vol) of ox gall (Sigma-Aldrich Bile bovine, CAS no. 8008-63-7). All cultures were incubated at 37°C for 0, 3, 6, 12, and 24 hours. Following incubation periods samples were serially diluted and plated in 3M Petrifilm aerobic count plates. As seen in the images below, total DE111 counts did not reduce in viability /concentration after contact with acidic buffers for 24 hours. Based on these results it was determined that DE111 is not sensitive to acid and is capable of maintaining viability in low pH concentrations.
Acid Survivability
Bacillus subtilis DE111 study 1
The objectives of this clinical study were to determine if daily consumption of Bacillus subtilis Strain DE111 (a spore-forming probiotic) at a 5 x 109 CFU/dose per day is safe for human consumption and efficacious as a probiotic. The tolerance and efficacy of encapsulated Bacillus subtilis Strain DE111 at a 5 x 109 CFU /dose per day was assessed in an average 20-day double-blind, randomized, placebo based study. Results showed that the majority of the blood parameters remained within normal ranges throughout; however, fasted serum glucose levels in the probiotic group (α≤0.05; P = 0.012) were significantly reduced. There were no significant differences presented in the average number of bowel movements per day within the probiotic group. There was a significant increase in the average number of bowel movements per day within the control group (α≤0.05; P=0.015). Significant differences in microbe colonization were present for B.subtilis and Bifidobacterium in the fecal colony counts. In conclusion, daily consumption of Bacillus subtilis Strain DE111 at a 5 x 109 CFU/dose per day can be recognized as a safe efficacious probiotic.
DE111 cardiovascular study 1
Cardiovascular disease (CVD) is the leading cause of death in the US and worldwide. By 2030 it is anticipated that CVD will claim the lives of more than 24 million people. Throughout the last decade, researchers have investigated the role of the gut microbiota in the development of CVD. Evidence exists for a positive correlation between Bifidobacterium and vascular function, glucose tolerance, and reduced systemic inflammation. Another probiotic species, Bacillus subtilis, has also been found to reduce cholesterol levels in human and animal models. In light of these data, we examined various measures of cardiovascular health after consumption of Bifidobacterium animalis subsp. Lactis strain BL04, with and without a cocktail of Escherichia coli-targeting bacteriophages (marketed as PreforPro), Bacillus subtilis strain DE111 or a maltodextrin-based placebo in a healthy human population. In a randomized, double-blind, placebo-controlled 4-week study conducted in individuals 18 to 65 years of age with a body mass index of 20 to 34.9, we saw no significant changes in measured CVD parameters among individuals consuming B. lactis with or without bacteriophages. However, 1 billion CFU/day B. subtilis supplementation resulted in a significant reduction in total cholesterol relative to baseline measures (-8 mg/dl; P=0.04, confidence interval (CI): -13.40, -0.19), as well as non-high-density lipoprotein-cholesterol (-11 mg/dl; P=0.01, CI: -12.43, -2.07). In addition we observed trending improvements in endothelial function (P=0.05, CI: -0.003, 0.370) and in low-density lipoprotein-cholesterol (P=0.06, CI:-12.29, 0.2864). Strikingly, these effects were seen in a largely healthy population. These data suggest that B. subtilis supplementation may be beneficial for improving risk factors associated with CVD. Further studies in populations of older adults or those with dyslipidaemia and endothelial dysfunction is warranted.
Bacillus subtills survival study 1
As part of an ongoing study to determine the true habitat of Bacillus species, we report here the isolation and characterisation of Bacillus subtilis from the human gastrointestinal tract (GIT). Strains were obtained from ileum biopsies as well as from faecal samples and their biotypes defined. 16S rRNA analysis revealed that most isolates of B. subtilis were highly conserved, in contrast to RAPD-PCR fingerprinting that showed greater diversity with 23 distinct RAPD types. The majority of B. subtilis strains examined possessed features that could be advantageous to survival within the GIT. This included the ability to form biofilms, to sporulate anaerobically and secretion of antimicrobials. At least one isolate was shown to form spores that carried an exosporium, a loosely attached outer layer to the mature endospore, this being the first report of B. subtilis spores carrying an exosporium. This study reinforces a growing view that B. subtilis and probably other species have adapted to life within the GIT and should be considered gut commensals rather than solely soil microorganisms.
DE111 children study 1
There is ample evidence suggesting that modulations in gut microbiota play an important role in inflammation and immunity. In particular, the microbiota of children is highly susceptible to environment influences, such as infections. Consequently, probiotics and their ability to promote and support a healthy microbiome have been increasingly studied. This study aimed at investigating the effects of a probiotic supplement (1 billion CFU/day Bacillus subtilis DE111) on the microbiome composition of preschool aged children attending day care. Healthy children aged 2-6 years old were randomized to receive either probiotic or placebo once a day for 8 weeks. No significant changes of the overall microbiome equilibrium were seen in between the two groups or from baseline to week 8. However, alpha diversity was increased in the probiotic group from baseline to week 8 (P<0.05), with no change in the placebo group. A decrease in the Firmicutes/Bacteroidetes ratio following probiotic supplementation (P<0.05) was also observed. Differential abundance analysis revealed an increase in Alistepes (P<0.01), Bacteroides (P<0.05), Parabacteroides (P<0.01), Odoribacter (P<0.001) and Rikenellaceae (P<0.001) in the probiotic group, most of which are involved in inflammation reduction. In addition, a decrease in Eisenbergiella (P<0.001), Lactobacillales (P<0.01) and Streptococcaceae (P<0.01), which is considered pro-inflammatory, were also observed in the probiotic group. Together with a reduction of the F/B ratio observed in the probiotic group, these results suggest probiotic supplementation with Bacillus subtilis DE111 introduce subtle but positive changes in the microbiome of children aged 2-6 years old.
DE111 children study 2
Children beginning preschool typically have an increased prevalence of gastrointestinal and respiratory infections. This study aimed to evaluate safety and efficacy of the probiotic Bacillus subtilis DE111® in gastrointestinal health and respiratory infections in preschool children. In a randomised, parallel, double-blind placebo-controlled study 102 day-care attending children aged 2-6 years received B. subtilis DE111® (1 × 109 CFU) or placebo once a day for 8 weeks. Participant diaries were completed by parents and evaluated by investigators to follow the incidence and duration of indicators of gastrointestinal health and respiratory infections as well as any adverse events. Saliva samples were collected at baseline and completion of the intervention to measure sIgA levels. A significant reduction in duration of vomiting (2 days vs. 14 days, p=0.045), duration of hard stools (0 days vs 15 days, p=0.044), and duration of overall gastrointestinal discomfort (18 days vs. 48 days, p=0.0499) was seen. No difference in incidence of respiratory infection was observed (41.3% probiotic vs 36.2% placebo, p=0.60). A statistically significant increase of sIgA levels was observed in the placebo group (1.37-fold, p<0.01), but not in the probiotic group (1.05-fold, p=0.61). Overall, data suggests intake of the probiotic B. subtilis DE111® is safe for use in children and supports a healthy gastrointestinal tract with a reduced duration of vomiting, hard stools and overall gastrointestinal discomfort.
FOS study 1
The use of probiotics and prebiotics has become firmly established due to their beneficial effects at the nutritional and therapeutic levels. Although their use is not new, the need to identify more effective species of probiotics which meet the criteria of stability, resistance and proliferation has gained considerable importance. Recognised species include Lactobacillus sporogenes, a spore-forming fermentative bacterium, that has demonstrated its utility and advantages in various studies and which also exhibits a high degree of safety. At the same time, not all prebiotics respond to the same bifidogenic parameters and they are the source of different nutrients which favour the colonic metabolism. Fructooligosaccharides (FOS) are noted for their considerable bifidogenic power and higher production of butyrate, a short-chain fatty acid which is essential to the colonocyte. The combination of both these effects has been defined as symbiotic and has generated significant interest with regard to their nutritional and therapeutic qualities in intestinal pathologies and in certain disorders of metabolism.
FOS study 2
Prebiotics, as currently conceived of, are all carbohydrates of relatively short chain length. To be effective they must reach the cecum. Present evidence concerning the 2 most studied prebiotics, fructooligosaccharides and inulin, is consistent with their resisting digestion by gastric acid and pancreatic enzymes in vivo. However, the wide variety of new candidate prebiotics becoming available for human use requires that a manageable set of in vitro tests be agreed on so that their nondigestibility and fermentability can be established without recourse to human studies in every case. In the large intestine, prebiotics, in addition to their selective effects on bifidobacteria and lactobacilli, influence many aspects of bowel function through fermentation. Short-chain fatty acids are a major product of prebiotic breakdown, but as yet, no characteristic pattern of fermentation acids has been identified. Through stimulation of bacterial growth and fermentation, prebiotics affect bowel habit and are mildly laxative. Perhaps more importantly, some are a potent source of hydrogen in the gut. Mild flatulence is frequently observed by subjects being fed prebiotics; in a significant number of subjects it is severe enough to be unacceptable and to discourage consumption. Prebiotics are like other carbohydrates that reach the cecum, such as nonstarch polysaccharides, sugar alcohols, and resistant starch, in being substrates for fermentation. They are, however, distinctive in their selective effect on the microflora and their propensity to produce flatulence.
Note: Although larger doses of FOS were used in the following four study summaries than are included in the current formulation, summaries are nonetheless included since they add weight to the totality of the research on FOS.
FOS study 3
Short-chain fructo-oligosaccharides (SC-FOS) are a mixture of oligosaccharides consisting of glucose linked to fructose units (Gfn; n = </= 4), which are not digested in the human small intestine but are fermented in the colon where they specifically promote the growth of bifidobacteria. In healthy volunteers, we assessed the tolerance and the threshold dose of SC-FOS that significantly increased fecal bifidobacteria counts and the possibility of a dose-response relationship. Randomly divided into five groups and eating their usual diets, healthy volunteers (40: 18 males, 22 females) ingested in two oral doses for 7 d a powder mixture containing (g SC-FOS/d): 0, G0; 2.5, G2.5; 5, G5; 10, G10; 20, G20. Stools were collected before (d1) and at the end (d8) of sugar consumption, and tolerance was evaluated using a daily chart. Total anaerobe counts were not affected by SC-FOS ingestion. Bifidobacteria counts at d8 were greater in groups G10 and G20 than in G0 and G2.5 (P < 0.05). Fecal pH did not differ among groups. A significant correlation between the dose of SC-FOS ingested and the fecal bifidobacteria counts was observed at d8 (r = 0.53; P < 0.01). Excess flatus was significantly more frequent in subjects consuming G20 than in those consuming G0, G2.5 or G5 (P < 0.05), and more intense in G20 than in G0 and G5 groups (P < 0.05). In conclusion, the optimal and well-tolerated dose of SC-FOS that significantly increased fecal bifidobacteria in healthy volunteers consuming their usual diet is 10 g/d.
FOS study 4
Five subjects completed the study consisting of a 30-d low fiber control period and another 30-d experimental period in which 10 g of FO was supplemented to a 3-d cycle menus. Status of large bowel function was monitored daily. Feces were completely collected on last 5 d of each period to determine the fecal characteristics and fecal short-chain fatty acid (SCFA) contents. Nutrient intake, body weight, arm anthropometric parameters, serum albumin, total protein, triglyceride, cholesterol, and electrolytes were measured as indices of nutritional status. Incorporation of FO significantly increased the defecation frequency, daily stool weight, weight per stool and decreased the use of enema. Concentrations of fecal acetate, propionate, i-butyrate, n-butyrate, i-valerate and n-valerate increased by 3–18 fold in the experimental period. Nutrient intake and anthropometric indices remained constant throughout the study. Serum concentrations of Na and K were decreased and concentrations of Ca and P were increased in the experimental period. Other blood biochemical parameters were not affected by the treatment. Therefore, we conclude that supplementation of FO was able to alleviate constipation, increase stool weight and fecal SCFA concentrations without affecting plasma lipid concentrations in the normolipidemic, constipated elderly men.
FOS study 5
The partial enzymatic hydrolysis of chicory inulin (GFn; 2 < or =n < or =60) yields an oligofructose preparation that is composed of both GFn-type and Fn-type oligosaccharides (2 < or =n < or =7; 2 < or =m < or =7), where G is glucose, F is fructose, and n is the number of beta(2–>1) bound fructose moieties. Human studies have shown that feeding GFn-type oligomers significantly modifies the composition of the fecal microflora especially by increasing the number of bifidobacteria. The experiments reported here were used to test the hypothesis that the Fn-type molecules have the same property. During a controlled feeding study, 8 volunteers (5 females and 3 males) consumed 8 g/d of an Fn-rich product for up to 5 wk. Fecal samples were collected and analyzed for total anaerobes, bifidobacteria, lactobacilli, bacteroides, coliforms and Clostridium perfringens. Both 2 and 5 wk of oligofructose feeding resulted in a selective increase in bifidobacteria (P<0.01). In addition, a daily intake of 8 g of the Fn-type oligofructose preparation reduced fecal pH and caused little intestinal discomfort.
FOS study 6
There is a need for studies on colonic fermentation in order to learn more about health and diseases of the colon. The aim of the present study was to evaluate the fate of two different doses of fructo-oligosaccharides (5 and 15 g/d) v. glucose in the intestine of healthy men. Twenty-four volunteers participated in a 5-week study. The study was a completely balanced multiple crossover trial using an orthogonal Latin-square design for three periods, with supplement periods of 7 d and two 7 d wash-out periods. Breath samples and faecal samples were collected. There was a clear gaseous response to the consumption of fructo-oligosaccharides. The highest dose significantly increased 24 h integrated excretion of breath H2 (P < 0.05). Breath H2 excretion after ingestion of 5 g fructo-oligosaccharides was higher than control, but did not reach significance. No effects on the total concentration of short-chain fatty acids in faeces were observed, no modification of the molar proportions of the various short-chain fatty acids was observed. The faecal pH did not change. No changes in faecal weight were observed. No fructo-oligosaccharides were recovered in faeces. We conclude that fructo-oligosaccharides added to the diet of young Western subjects are fully metabolized in the large intestine. The level of fermentation seem to be dose-dependent.
- References
- Hong HA, Khaneja R, Tam NMK, et al. Bacillus subtilis isolated from the human gastrointestinal tract. Research in Microbiology. 2008; 160(2):134-43.
- DE111® pH Survivability. Deerland Probiotics & Enzymes. May 26, 2017. Not published.
- Labellarte GM, Maher M, Healey A, Deaton J. Tolerance and efficacy of the probiotic DE111® delivered in capsule form. Department of Biology, University of Wisconsin-La Crosse. Unpublished. 2015:22 pgs.
- Trotter RE, Vazquez AR, Grubb DS, et al. Bacillus subtilis DE111 intake may improve blood lipids and endothelial function in healthy adults. Benef Microbes. 2020 Nov 15;11(7):621-630.
- Hong HA, Khaneja R, Tam NMK, et al. Bacillus subtilis isolated from the human gastrointestinal tract. Research in Microbiology. 2008; 160(2):134-43.
- Paytuví-Gallart A, Sanseverino W, Winger AM. Daily intake of probiotic strain Bacillus subtilis DE111 supports a healthy microbiome in children attending day-care. Benef Microbes. 2020 Nov 15;11(7):611-620.
- Slivnik M, Kristan KC, Lipovec NC, Locatelli I, Orel R, Winger AM. Effect of Daily Bacillus subtilis DE111® Intake on Gastrointestinal Health and Respiratory Infections in Children Attending Day-care: A Randomised, Parallel, Double-blind, Placebo-controlled Study. J Prob Health. 2020; 8:225.
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