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LactoStim® Prebiotic Studies

Most prebiotics are types of soluble fiber that are not digested by mammalian digestive systems but, to varying degrees, are digested by certain intestinal bacteria. Fructooligosaccharides (FOS) and inulin are examples of such prebiotics. The best fiber-based prebiotics are composed of oligosacchirdes that are selective to probiotics in the Lactobacillus and Bifidobacteria genera.

All fiber-based prebiotics have two things in common:

  1. They need to be consumed at 3-8 grams per day to be effective
  2. Probiotics grow without them but show enhanced growth in their presence

LactoStim® is a completely new type of prebiotic that does not contain fiber. Instead, we use a combination of  sunflower lecithin and oleic acid which show dramatic probiotic stimulation in milligram quantities (10-500 mg daily) unlike comparable, fiber-based prebiotics. Not only does LactoStim stimulate the growth of encapsulated probiotics, but it stimulates the growth of all probiotic bacteria by aiding the membrane permeability of probiotic strains permitting enhanced bio-availabilty of nutrients critical to the healthy growth and colonization of probiotic bacteria. This puts probiotic bacteria on a level playing field with other intestinal microbes that do not require additional supplementation.

LactoStim is an all natural biosurfactant that replaces polysorbate 80 as a bacterial membrane activator in lactic acid bacteria such as Lactobacillus and Bifidobacterium probiotics. Polysorbate 80 is found in bacteriological growth media used for cultivating probiotics — MRS and LBS probiotic growth media are two typical examples. When polysorbate 80 is omitted from otherwise nutrient-rich media, little or no probiotic growth will be observed.  

The bacterial membrane exists inside the cell wall and acts as a gate that allows nutrients in and waste matter out of the cytoplasm. An activated membrane allows for more efficient transportation in both directions and thus increases both bacterial growth and metabolic activity. Most intestinal bacteria do not require membrane activation so they do not benefit from polysorbate 80 nor LactoStim. This is good news as it makes LactoStim selective for probiotic bacteria.

In the following experiments, we measured the growth and activity of our supplements as well multiple competitors' products both in the presence of LactoStim and without. We used a modified MRS broth with the recipe provided below.

The results show how effective LactoStim is at improving the growth and activity of not only our own probiotic supplements but our competitors as well.

Experiment: LactoStim Prebiotic Growth & Activity

Purpose:

To study the growth and lactic acid production of various probiotic strains using LactoStim as a substitute for polysorbate 80 in a standard Difco MRS Lactobacillus broth. Various commercial probiotics were inoculated into this broth at 0.1% and incubated at 37°C for 12 hours. Growth, measured as turbidity, and activity, measured as percent of lactic acid produced, were recorded at 2 hour intervals for the duration of the experiment.

Difco Lactobacillus MRS Broth Formula (per liter)
IngredientAmount
Proteose Peptone No. 3 10.0 grams
Beef Extract 10.0 grams
Yeast Extract 5.0 grams
Dextrose 20.0 grams
Polysorbate 80 1.0 grams
Ammonium Citrate 2.0 grams
Sodium Acetate 5.0 grams
Magnesium Sulfate 0.1 grams
Manganese Sulfate 0.05 grams
Dipotassium Phosphate 2.0 grams
Distilled Water 1 liter

Procedure:

  1. Prepare 400 ml flasks of MRS broth using the above formula as a positive control; the above formula sans polysorbate 80 as a negative control; and the above formula substituting LactoStim for polysorbate 80 in an equal amount for each test. 
  2. Sterilize at 121°C for 15 minutes in an autoclave. 
  3. Temper each flask to 47°C. 
  4. Aseptically inoculate each flask with 0.14 grams of individual probiotic strain to be tested. 
  5. At each time interval, aseptically transfer a 30 ml sample from each flask onto a HACH 2100N Turbidimeter sample cell. 
  6. Read turbidity of each sample and report the results in NTU's (Nephelometric Turbidity Units). 
  7. Transfer the same 30 ml sample used for the turbidity reading into a 250 ml glass beaker. 
  8. Record pH. 
  9. Titrate sample using 0.1N NaOH to an endpoint of pH 6.8. 
  10. Record mls of 0.1N NaOH used. 
  11. Calculate % Lactic Acid production using the following formula:

% Lactic Acid = (((mls of 0.1N NaOH) x (0.1N) x (90 gm/mol))/mls of sample) x (1 L/1000 ml) x 100

% Lactic Acid = ((mls of 0.1N NaOH) x (0.9))/30 

Results:

LactoStim Growth Study: Theralac®

Chart depicting growth rate of Theralac with and without Lactostim

30 billion CFU/capsule
(~0.45 gm/capsule)

Lactobacillus paracasei (F-19)

Lactobacillus rhamnosus (LR-44)

Lactobacillus acidophilus (LA-1)

Bifidobacterium lactis (BL-34)

Bifidobacterium lactis (Bi-07)

5 billion

2 billion

10 billion

10 billion

3 billion

Chart depicting Lactic Acid Activity in Theralac with and without LactoStim

Other Ingredients

Cellulose, Sodium Alginate, Sodium Phosphate, LactoStim (Lecithin + Oleic Acid), Hydroxypropyl Methylcellulose (HPMC vegetable capsule), Silica, Magnesium Stearate

LactoStim Growth Study: Competitor A

Chart depicting growth rate of competitor a with and without Lactostim

15 billion CFU/gram
(~0.45 gm/capsule)

Lactobacillus acidophilus

Lactobacillus rhamnosus

Streptococcus thermophilus

Lactobacillus plantarum

Bifidobacterium bifidum

Lactobacillus bulgaricus

Bifidobacterium longum

Lactobacillus salivarius

45%

25%

10%

7%

6%

3%

3%

1%

Chart depicting Lactic Acid Activity in a competitor with and without LactoStim

Other Ingredients

Microcrystalline cellulose, magnesium stearate, ascorbic acid, vegetarian capsule (water and hypromellose)

LactoStim Growth Study: Competitor B

Chart depicting growth rate of competitor b with and without Lactostim

10 billion CFU/capsule
(~0.355 gm/capsule)

Lactobacillus GG

10 billion

Chart depicting Lactic Acid Activity in competitor b with and without LactoStim

Other Ingredients

Inulin (245 mg), gelatin (capsule)