August 15, 2023

Important Takeaways from the Latest Science on Spore-Based Probiotics

In the Dirt, in Our Water
– Our History With Spores

If you’re a pet owner or parent, you might be familiar with your fur baby or kid eating mouthfuls of dirt. Although these might incite momentary panic, perhaps there is something to this innate urge to consume soil…

Let’s take a deeper look!

Spore-based probiotics or spore-based organisms (SBO) are tiny organisms produced by bacteria naturally occurring in the soil. Before the modernization of farming and sterilization techniques, we had daily exposure to SBOs found on our produce or through drinking fresh spring or well water.¹

These spores have two distinct morphologies – endospore and vegetative cell. The resilience of SBOs lies in their ability to produce endospores. Endospores are the most durable type of cells found in nature and represent a survival strategy of bacteria. Bacillus spores have been isolated from amber that is more than 25 million years old, demonstrating the metabolic dormancy and protective capability of spores.² However, to provide benefits to their hosts, the spore-based probiotics must be metabolically active in their vegetative state.

The Role of Endospores in Probiotic Resilience

The outer surface of endospores has hydrophobic characteristics making the spores insoluble. Germination happens when nutrients or other agents such as lysozyme, salts, high pressures, and cationic surfactants bind to germinant receptors on the inner membranes that trigger a cascade of events leading to water entering the spore cores.² This rehydration activates spore enzymes that hydrolyze the spore cortex, allowing the spores to start germination. When it comes to their journey through the GI tract, the low pH of the stomach activates the spores before they reach the nutrient-rich small intestine, where they proliferate.

The endospores that encapsulate the probiotic strains make them resilient to stomach and bile acid as well as resistant to heat, light, ultraviolet radiation, solvents, and hydrogen peroxide.^² This allows for germination in the small intestine, the optimal functional location for probiotic residence. To demonstrate their ability to reach the intestines, a randomized, double-blind, placebo-controlled study involving 11 participants with ileostomies showed the Bacillus subtilis DE111 spores and vegetative cells present in the ileum effluent just three hours after ingestion. Six hours after ingestion, the spore and vegetative concentration increased and remained constant to the final time point of 8h.³

Traditional Probiotics vs. Spore-based: What’s the Difference?

There is a huge distinction between spore-based probiotics that are shelf-stable and traditional multi-strain probiotics that need to be refrigerated.

The journey for oral probiotics is a strenuous one that greatly affects the ability of traditional probiotics to provide benefits to the host. Probiotics must endure gastric acid, bile salts, and degrading enzymes, all before they arrive at their functional site in the gastrointestinal tract. Just in the stomach alone, due to ionic strength, enzyme activity, and mechanical churning, the viable cells of specific probiotics such as Bifidobacterium longum and Bifidobacterium brevehave been shown to be undetectable in simulated gastric juice within an hour.If they make it to the colon, viable probiotics must then manage to colonize the gut in competition with indigenous bacteria.

As noted above, spore-based probiotics have the ability to travel successfully through the gastrointestinal system and germinate in the small intestine. What’s even more profound is that, unlike traditional probiotics that reproduce to form a stable replicating population, spores can recondition the gut through microbial diversity and support the growth of beneficial bacteria, including keystone species such as Akkermansia.^¹

**Let’s Take a Closer Look at a Few Bacillus Species:**

Bacillus Species – A Highly Researched SBO

Bacillus species are a group of saprophytic Gram-positive, rod-shaped, spore-forming aerobic or facultatively anaerobic bacteria.⁵ Traditionally Bacillus species have been used for the production and preservation of food in many different countries and, more recently, have been incorporated into functional foods. The genus Bacillus is closely related to Lactobacillus spp., which both belong to the phylum Firmicutes. Of the hundred species contained in the Bacillus genus, only a select few are used as probiotics, including Bacillus subtilis, Bacillus coagulans, Bacillus clausii, Bacillus pumilus, Bacillus licheniformis, Bacillus natto (subtilis), Bacillus polyfermentans, Bacillus cereus var. toyoi, and Bacillus cereus.¹¹

Although probiotics are commonly used for gastrointestinal health, spores provide astonishing multisystem health benefits. Besides supporting a diverse microbiome, nutrient absorption, reducing digestive discomfort such as gas and bloating, and regulating bowel function, Bacillus species play a role in modulating a healthy inflammatory response, supporting immune regulation, hormonal balance, and even cardiovascular health.

Bacillus Coagulans MTCC 5856 (LactoSpore)

Bacillus coagulans MTCC 5856, or Lactospore, is a lactic acid-producing bacteria, hence its name. The production of levorotatory L(+)lactic acid (the form most readily metabolized in glycogen synthesis, thus not contributing to metabolic acidosis) allows this spore to change the luminal pH, which inhibits the growth of pathogens in the intestines while supporting an environment favorable to obligate microorganisms. Additionally, this spore produces otherbacteriocins, such as coagulin, as well as substances, such as short-chain fatty acids (SCFAs), that are critical for the health of the colon.⁷

B. coagulans MTCC 5856 has been studied to be safe for human consumption and is eliminated from the intestinal tract within four to five days unless administration is continued. Human clinical trials on this spore show therapeutic benefits in various gastrointestinal conditions such as chronic and acute gastroenteritis, diarrhea, constipation, colic in babies, diarrhea in newborns, IBS, IBD, and digestive discomfort such as flatulence, indigestion, cramps, heartburn, and bloating.

In one study, Lactospore demonstrated a significant decrease in gastrointestinal symptoms such as bloating, vomiting, diarrhea, abdominal pain, and stool frequency in individuals with diarrhea-predominant IBS.⁸

Another randomized clinical trial conducted amongst 40 participants with IBS and major depressive disorder found that supplementation with 2 billion CFUs of Bacillus coagulans MTCC 5856 for 90 days not only significantly improved IBS symptoms related to quality of life (IBS-QOL) but also led to improvement in several validated scales which assess mood, including HAM-D (Hamilton Rating Scale for Depression), MADRS (Montgomery–Åsberg Depression Rating Scale), and CES-D (Centre for Epidemiological Studies–Depression Scale) compared to placebo.⁹

Besides its support to the gastrointestinal tract, Bacillus coagulans has been shown to have anti-inflammatory effects through cytokine modulation, inhibition of reactive oxygen species, and enhanced phagocytosis.⁶ It also has high levels of b-galactosidase (lactase) activity in vitro and may also have lactic acid dehydrogenase activity, thereby enhancing the digestibility of lactose in those who are lactose intolerant.⁷ Other benefits have been seen when used for non-specific vaginitis, UTIs, immunodeficiency, and lipid lower effects.

B. coagulans daily doses reported in peer-reviewed research range from 100 million to 5 billion CFUs. Beneficial effects were noted in earlier studies, even at dosages as low as 100 million CFUs daily. Currently, a typical dosage recommendation is 1.5 billion colony-forming units 2-3 times daily.⁷

Bacillus Subtilis DE111

Bacillus subtilis DE111 is a clinically tested strain that supports digestion and immune health. Traditionally used in Japan and Africa in the preparation of fermented foods, Bacillus subtilis can produce a variety of different enzymes, including lipase, carbohydrases, amylase, protease, and cellulase, and works with endogenous enzymes to support the absorption of nutrients.¹⁰ Additionally, DE111 produces specific proteases that can help break down gluten and other allergen foods such as egg, soy, and dairy to reduce the effects of sensitivity in those who react to them.

Genome analysis of DE111 has revealed the presence of de novo synthesis of water and fat-soluble vitamins such as B1 (Thiamine), B2 (Riboflavin), B6 (Pyridoxine), B7 (Biotin), B9 (Folate), and vitamin K2 (Menaquinone) and amino acids including five essential amino acids (threonine, tryptophan, methionine, leucine, and lysine) and two non-essential amino acids (cysteine and arginine).¹¹

Recent evidence suggests that Bacillus subtilis may be part of normal gut microbiota and play a critical role in immune development and GI health. It not only completes its life cycle in the GI tract, but evidence of B. subtilis isolated from the GI tract shows faster germination than strains produced in a lab setting.¹² It is hypothesized that B. subtilis can create its own aerobic environment in the gut by producing exopolysaccharide (EPS) biofilm, which can also be used as a prebiotic to stimulate the growth of other beneficial flora.¹²

With the ability to survive in aerobic and anaerobic conditions, utilize a variety of growth nutrients, and germinate quickly, Bacillus subtilis provides a range of benefits, including antimicrobial, antidiarrheal, immunostimulatory, competitive exclusion of pathogens, prevention of intestinal inflammation, and normalization of intestinal flora.⁶ Additionally, this strain has been clinically proven to support cardiovascular health, infant and children’s gastrointestinal and immune health, as well as support cortisol levels and enhance athletic recovery.¹³

Below is a chart showing the vast amounts of antimicrobial compounds Bacillus subtilis can produce, contributing to this strain's ability to address a broad spectrum of pathogens.

Several double-blind, randomized, placebo-controlled trials have been conducted using DE111. One such study involving 44 healthy individuals who supplemented with the strain for 30 days found it supported microbial population and digestion, suppressed intestinal and systemic inflammation, and reduced total and non-HDL cholesterol, glucose, and triglyceride levels.¹⁴ DE111 helps to reduce blood sugar spikes by converting sugars and fats to SCFAs, notably butyrate. It also improved reactive hyperemia index through improved elasticity of blood vessels. This same study also showed immune-modulatory and anti-inflammatory effects on regulatory T cells, increased CD25+ and CD25+FoxP3+ Tregs, as well as CD4+CD8+ double-positive T cells in response to stimulation with lipopolysaccharide (LPS, or endotoxin).¹⁴

This research demonstrates DE111’s ability to play a role in helping its host combat the inflammatory response induced by low-grade LPS exposure commonly associated with metabolic endotoxemia. It allows the host's immune system to respond faster to threats and return to baseline more quickly.

In another study involving 50 individuals with occasional constipation and/or diarrhea, DE111 was supplemented for 90 days and was found to help protect against occasional constipation or diarrhea and support overall regularity, transit, and composition of fecal matter.¹⁵

In adult athletes, DE111 reduced circulating levels of TNF-alpha with a trend of lowering cortisol.¹⁶ What’s more is that female athletes saw improved lower body strength, body composition, exercise performance, and decreased body fat when given for ten weeks during offseason training.¹⁷

Finally, this strain has been well-documented to be beneficial for supporting the gastrointestinal and immune health of infants and children. One study with 91 children ages 2–6 used DE111 for eight weeks and saw an increase in alpha diversity of the microbiome with favorable changes, including a reduction in Firmicutes/Bacteroidetes ratio.¹⁸ It was also shown to reduce the duration of vomiting and GI discomfort (bloating, diarrhea, constipation) by 62% compared to placebo.¹⁹

Regarding dosing, one study showed daily ingestion of one capsule containing approximately 5 x 10⁹ or 5 million colony forming units (CFU) per capsule of B. subtilis was well tolerated in healthy young adults consuming their usual and variable diets, as reflected by blood levels of important biomarkers. Markers of systemic acceptance, such as CRP and liver enzymes, remained within acceptable ranges, and gastrointestinal symptoms and bowel habits, if anything, improved with probiotic capsule consumption.²⁰

Bacillus Clausii SC108 (Munispore)

Bacillus clausii SC108 naturally occurs in whole grains and in fermented foods and beverages. Although transient in nature, it has the ability to adhere to the epithelial lining of the gut using adhesion-related proteins, which permits it to increase germination in the GI tract before it re-sporulates.²¹ Once present in the gut, it crowds out pathogenic microbes allowing healthy gut flora to thrive.

Bacillus clausii stands out from other Bacillus strains with its ability to modulate the immune response, as shown in vitro and in vivo. One such study showed this strain was able to lower E. coli lipopolysaccharide-induced proinflammatory cytokines significantly.²⁰ Besides immune modulation, Munispore also has immunostimulatory properties allowing for quicker and more efficient immune cell response before returning to baseline.

Bacillus clausii is characterized by intrinsic resistance to penicillins, cephalosporins, aminoglycosides, and the macrolides. Bacillus clausii also possesses an acquired resistance to tetracycline, chloramphenicol, and to rifampicin and thus is safe to use in conjunction with antibiotic therapy.²²

Additionally, this strain has its own antimicrobial effects. One study showed the vegetative state of Bacillus clausii SC108 reduced the growth of E. coli, S. aureus, and P. aeruginosa in vitro.²⁰

Further activities of this strain include its ability to modulate the activity of genes linked to inflammatory, immune, and defense responses, as well as to intestinal permeability, cell adhesion, cell growth, cell differentiation, cell signaling, apoptosis, signal transcription, and transduction.⁶

Munispore's robust antioxidant profile contributes to its ability to combat oxidative stress, minimize cell damage, and support healing and immune regulation. Specifically, this antioxidant capacity has been shown to protect against free radicals that affect tight junctions and help repair those junctions more quickly. Although a handful of probiotics exhibit antioxidant activity, Munispore’s total antioxidant capacity (TAC) is significantly higher than the well-known Lactobacillus rhamnosus when it comes to catalase activity.²⁰ Additionally, this strain has B-galactosidase activity and the capacity to synthesize amino acids and B vitamins, notably pantothenic acid and cobalamin.²⁰

As for its specific support to gastrointestinal health, B. clausii has long been used in Italy to address viral diarrhea in children and antibiotic-related GI side effects. One study found that Bacillus clausii was able to produce a protease that reduces the cytotoxic effects of Clostridium difficile and Bacillus cereus, which researchers suspect explains its benefits against antibiotic-associated diarrhea.²³

This strain has also been used as an adjunct therapy in the treatment of Helicobacter pylori, as it was shown to alleviate symptoms of acute diarrhea, nausea, and epigastric pain that may come from anti-H. Pyloriantibiotic treatment. A few studies also demonstrate its ability to alleviate symptoms of acute diarrhea (duration, frequency, abdominal pain, stool consistency) in adults and children.^{25, 26} Bacillus clausii shows promising support for addressing small intestinal bacterial overgrowth (SIBO), a common etiology for IBS. Supplementation of the strain over one month resulted in the normalization of hydrogen glucose breath test in 47% of participants, comparable to the success rate of many single broad-spectrum antibiotics.²⁷

In one study, patients aged 6-17 years with IBS (Rome IV) for at least 2 months were randomized to receive either B. clausii (oral suspension, total dose 4 billion spores/day) or placebo once daily for 8 weeks. All patients also received conventional treatment. The primary endpoint was the difference in the proportion of patients with clinical improvements at Week 8. 73.6 % of those receiving B. Clausii compared to 78.5% of those receiving the placebo saw symptom improvement demonstrating the safety and efficiency of this spore at this dose.²⁸

Conclusion:

Once commonly part of our daily exposure, spore-based probiotics provide a safe and effective way to support our overall health. With their resilient nature to survive the treacherous journey to the intestines and implications that reach far outside the walls of the gastrointestinal tract, these microorganisms should be reconsidered as part of our daily self-care regimens. Indeed, they can play an impactful and therapeutic role in addressing digestive health, immune regulation, inflammatory control, and supporting cardiovascular and hormonal health.

Multistrain traditional probiotics, although not as stable as SBOs, still have useful applications when it comes to enhancing the diversity of our patients' microbiomes and, when used in conjunction with spore-based probiotics, may offer both immediate and long-lasting therapeutic change.

Learn more about spore-based probiotics.

Resources:

Professional Benefits

Biocidin Academy

Important Takeaways from the Latest Science on Spore-Based Probiotics

In the Dirt, in Our Water
– Our History With Spores

The Role of Endospores in Probiotic Resilience

Traditional Probiotics vs. Spore-based: What’s the Difference?

**Let’s Take a Closer Look at a Few Bacillus Species:**

Bacillus Species – A Highly Researched SBO

Bacillus Coagulans MTCC 5856 (LactoSpore)

Bacillus Subtilis DE111

Bacillus Clausii SC108 (Munispore)

Conclusion:

Biocidin Academy

In the Dirt, in Our Water – Our History With Spores

The Role of Endospores in Probiotic Resilience

Traditional Probiotics vs. Spore-based: What’s the Difference?

Let’s Take a Closer Look at a Few Bacillus Species:

Bacillus Species – A Highly Researched SBO

Bacillus Coagulans MTCC 5856 (LactoSpore)

Bacillus Subtilis DE111

Bacillus Clausii SC108 (Munispore)

Conclusion:

In the Dirt, in Our Water
– Our History With Spores

**Let’s Take a Closer Look at a Few Bacillus Species:**