Gut health has an importance that goes beyond the digestive and assimilation processes carried out on it. There is more and more evidence of its importance in the development of diseases that apparently are far from what happens in the intestine. Given the importance of proper intestinal health for the maintenance of general health, the gut is also known as the second brain.

In our gut cohabits a bacterial flora that is known as microbiota. We have more than 100 trillion bacteria (1×10¹⁴), which are not equally distributed throughout the gastrointestinal tract. They pass from 10 to 1000 bacteria per gram in the stomach to more than 1 trillion per gram in the intestine (1×10¹²). In fact, we have 10 times more bacteria than cells in our body. In addition, these bacteria have their own genes that represent 100 times the total human genes. The term microbioma refers to this set of genes present in the microorganisms that inhabit our gastrointestinal tract.

Despite bacterial flora is specific to each person, it can be said that these bacteria are distributed in 150 species of the more than 1000 species that may be present. This bacterial diversity is important for our health.

Increase in the number of bacteria along the intestinal tract

Under normal conditions, the bacteria are placed on the mucosa of the intestinal epithelium and does not penetrate into the body. However, contrary to what most people think, the gut microflora has an influence that extends beyond the intestinal lumen.

The intestinal microflora acts on three levels:

• Level 1: Bacteria – bacteria interaction
• Level 2: Bacteria – intestinal mucosa interaction
• Level 3: Bacteria – host immune system interaction.

At the first level, bacteria interact with other bacteria, either beneficial as pathogenic. At this level, the beneficial bacteria and commensal establish symbiotic relationships with other, but also release important substances such as lactic acid and bacteriocins that allow them to control and compete with the pathogenic flora.

At the second level, the microflora initiates the interaction with the host throughout the intestinal microvilli. In this sense, bacteria establish recognition between the lipopolysaccharides present in the bacterial wall with the receptors present in the membrane of the enterocytes. This recognition is very important because it allows the creation of beneficial interactions with immune intestinal cells and allows the maintenance of mucosa integrity.

At the third level, bacteria contact with the host immune system, thanks to the presence of the dendritic cells located in the basal part of the intestinal mucosa. Dendritic cells are the most powerful antigen presenting cells and control the type of immune response to be established (pro-inflammatory or regulatory), as well as its location after making antigen presentation.

Dendritic cells are, therefore, very efficient in controlling the delicate balance between tolerance and immunity in an environment exposed to a huge quantity of antigens as intestines are. Any factor affecting dendritic cells activity can have an impact on its functionality, even in the development of intestinal pathologies like celiac disease or inflammatory bowel disease.

Several factors can alter the presence, concentration, distribution and type of bacteria that inhabit our intestine. Situations such as malnutrition or unbalanced nutrition, antibiotic consumption or stress can lead to an imbalanced flora situation known as dysbiosis that facilitates the onset of diseases.

On the left an intestinal mucosa with proper bacterial flora and on the right an intestinal mucosa suffering dysbiosis.

Both, the World Health Organization (WHO) and the Food and Agriculture Organization of the United Nations (FAO), recognize that the alteration of the intestinal flora leads to the occurrence of diseases that are directly related to the gut, but also diseases or conditions indirectly related to it:

The WHO (World Health Organization) and the FAO (Food and Agriculture Organization of the United Nations) recommend the use of probiotics for preventing and treating disorders caused by an unbalanced microflora and advise the medical community to consider this application for preventive and therapeutic actions.

It is worth remembering what a probiotic is since, recently, it has been introduced in the food supplements market products that are sold as probiotics, which do not meet the definition given by WHO:

“A probiotic is a living microorganism which, when administered in adequate amounts, confers health benefits to the host”.

In order to classify bacteria as probiotic, it is necessary to provide enough documented scientific evidence on the beneficial health effects because not all bacteria have the same effectiveness, even belonging to the same genus and species. Hence, it is important a perfect characterization of bacterial strains, which allows a correct selection of probiotics.

Let’s take as an example a study on the degree of inhibition of Candida albicans by different individual strains of Bifidobacterium, Lactobacillus, Enterococcus and Lactococcus.

Percentage of inhibition of Candida albicans by different bacterial species and strains properly characterized.

The figure shows that of the 3 strains Bifidobacterium studied (two of B. bifidum and one of B. lactis), only the two strains of B. bifidum presented an inhibition of more than 30% of Candida, but only the strain characterized as B. bifidum W28 inhibited Candida more than 80% when compared to B. bifidum W23 strain.

This study demonstrates also that of the three characterized strains of Lactobacillus acidophilus used (W22, W55 and W74), the strain L. acidophilus W55 did not inhibit the growth of Candida, unlike the characterized strains W22 and W74 did.

Therefore, the characterization allows to highlight the properties of a bacteria, showing that the properties of bacteria X are different from bacteria Y. In addition, the use of well-characterized strains makes that the combination of different bacteria with different properties increase their effectiveness.

It conclusion, multi-strain and multispecies mixtures are much more beneficial than the use of individual bacteria, as these mixtures will act on the three levels of the intestine.

10 years ago, Solchem Nature S.L. positioned a multispecies and multi-strain probiotic mixture in the market specifically selected for overall health and well-being, which has become one of the best mixtures for different gastrointestinal situations and general health, thanks to its unique properties that clearly distinguish it from any other similar mixture.

In recent years, there has been a continuous research in the field of probiotics about the relationships among gut microbiota, probiotics, prebiotics, health and disease, with the objective to improve Solchem’s multispecies and multi-strain probiotic mixture.

With the objective of increasing the effectiveness of the probiotic-prebiotic mixture, it has been developed a specific technology resulting in a new and improved formulation.

Research has led to an improvement of the existing mixture, which is presented under the name Megaflora 9 evo. This mixture is based in an improved prebiotic matrix containing ingredients specifically selected for their efficacy on the different Megaflora 9 evo bacterial strains, in order they can act at three intestinal levels.

Fruit of an innovative research supporting probiotic properties; SOLCHEM NATURE S.L. presents a specific matrix that includes prebiotic ingredients, minerals and enzymes.

The balance of these ingredients in the matrix has been widely studied and the amounts of each ingredient carefully adjusted to suit the needs of the 9 bacterial strains of Megaflora 9 evo. The result is a highly active multispecies probiotics.

Specific prebiotic ingredients for bacterial strains

Prebiotics are food components, primarily carbohydrates and fibre that selectively promotes growth and/or activity of bacteria in the large intestine. Research has shown that each probiotic strain has a specific preference for one type of prebiotic, as shown for example for L. acidophilus.

Growth of different strains of Lactobacillus acidophilus in different prebiotics substrates.
Abbreviations: Polydex (Polydextrose), GOS (Galacto-oligosaccharides), scFOS (Fructo-oligosaccharides short chain)

Vegetable protein

The specific selected ingredients that are part of the matrix makes bacteria of Megaflora 9 evo resistant to the gastric juice action and helps bacteria to reach the intestine in an active state. In this state, bacteria are able to produce lactic acid, short-chain fatty acids and other metabolites.

The matrix provides the nutritional support that bacteria needs to colonize and multiply themselves in the gut, resulting in an even more effective probiotic product.

Among the different ingredients, the presence of a vegetable protein as a prebiotic ingredient in the matrix produces a positive effect on the metabolic activity of the bacteria, mainly in the production of lactic acid.

Lactic acid producing activity favoured by the matrix.

Furthermore, the presence of the vegetable protein facilitates and shortens the rehydration of the product. Ussually, the rehydration of probiotics can take up to 10 minutes, whereas the presence of this vegetable protein shortens it to 1 minute.

In practice, this means that the probiotic formulation may be consumed almost immediately after adding water. The vegetable protein reduces the rehydration time at the same time that shows the highest production of lactic acid.

Impact of the incorporation of the vegetable protein in prebiotic and mineral matrix when rehydrating Megaflora 9 evo.

In summary, the specific technology used in Megaflora 9 evo production includes a special matrix that protects and nourishes the bacteria. These ingredients specifically selected and tested have shown that:

• Increase the product shelf life.
• Increase the gastrointestinal bacteria survival.
• Increase the metabolic activity of lactic acid producing bacteria in the gut.

If something characterizes Megaflora 9 evo as a new generation of probiotics is not only the presence of a special matrix, as has been explained, but a careful selection of bacterial strains.

Megaflora 9 evo contains nine bacterial strains well characterized comprised in 4 different species with a concentration of 2.000.000.000 cfu/g:

• Bifidobacterium lactis W51
• Bifidobacterium lactis W52
• Enterococcus faecium W54
• Lactobacillus acidophilus W22
• Lactobacillus paracasei W20
• Lactobacillus plantarum W1
• Lactobacillus plantarum W21
• Lactobacillus salivarius W24
• Lactococcus lactis W19

These nine strains are integrated into the matrix and have shown efficacy in three levels:

• Level 1: Inhibition of pathogens.
• Level 2: keeping barrier function.
• Level 3: stimulate the production of immunomodulatory cytokines

>> Action at Level 1: Inhibition of pathogenic

In traditional probiotic products, a large number of viable cells die during exposure to environmental conditions. Megaflora 9 evo matrix protects the bacteria against unfavourable conditions and allows to store the product at room temperature. The matrix significantly improves the stability giving a long shelf life.

The effectiveness of a probiotic product depends on the total viable bacterial count in the product. For this reason, the half-life of a probiotic products is based on the total of viable cells during storage time. The best method to measure the half-life of the product is by a real time stability test (RTS), where the bacterial count is expressed as colonies forming units (cfu) per gram, meaning cfu as living and viable bacteria.

The recent development of Megaflora 9 evo makes not possible to have a real shelf life under RTS test. However, it was carried out an accelerated stability test at 37° C ± 2° C for 4 weeks. This study considered the maximum temperature at which no bacterial damage occurs, but allows to check the stability in a short period that can be extrapolated to a situation of 25ºC stability.

Megaflora 9 evo samples were tested at T0 to T2, T3 and T4 times. The result shows that the concentration of bacteria was stable during all the time with a concentration that exceeded one billion per gram, which is the guaranteed concentration at the end of the shelf life of the product.

Megaflora 9 evo has the same stable behaviour than its probiotic and prebiotic predecessor, which has a stability of 48 months in RTS test.

The accelerated stability study has demonstrated that Megaflora 9 evo has a comparable stability than its predecessor and, therefore, at room temperature the same behaviour is expected. To prove this, a RTS test on Megaflora 9 evo is on going.

Accelerated stability test at 36 ° C for 4 weeks, of Megaflora 9 evo.

Surviving the path of the gastrointestinal tract is the first basic function of a probiotic based product. The more bacteria survive the passage through the gastrointestinal tract, better will be the effect on our body.

When bacteria are unprotected, a large number of viable cells is destroyed as a result of the low pH of the stomach, enzymes and digestive fluids.

In traditional products based on probiotics, a large quantity of bacteria are destroyed by the fluids of the GI tract. It is estimated that the survival of the bacteria that are ingested without any protection ranges from 0.5 to 1%, when they reach the intestine, meaning that, for example, from a concentration of 100,000 million bacteria per gram of product supplied, only 1,000 million bacteria will reach the intestine alive.

Probiotic blend: with 100.000 million bacteria/g (1 x 10¹¹)
1% de survival = 1.000 million viable bacteria (1 x 10⁹)

Megaflora 9 evo: with 2.000 million bacteria/g (2 x 10⁹)
minimum 90% survival = more than 1.800 million viable bacteria (>1,8 x 10⁹)

The technology used in Megaflora 9 evo production protects the bacteria against the acidic conditions of the stomach and keeps bacteria alive. This technology ensures a survival of more than the 90% of the initial concentration. This explains why the concentration of bacteria in Megaflora 9 evo does not need to be as high as other commercial preparations, in which an important part of the bacteria suffer a high mortality during their transit through the GI passage.

In vitro model that simulates the digestive system showing compartmentalisation, pH, enzyme and bile secretion over 6 hours digestion.

Gastrointestinal survival can be studied on a in vitro model developed by the University of Maastricht. This model simulates the passage of bacteria through the GI tract and allows to calculate the survival of the strains, and its resistance to gastric acid, pepsin, pancreatin and bile salts.

The model imitates the compartmentalization and timing of transit in every part of the GI tract, regulating the secretion of pH, addition of enzymes and secretion of bile onto the sample to be studied.

The experiment is run at 37 ° C and the total count of bacteria in the sample is measured at four different times:

• After rehydration of the sample (T = ‘Ah)
• After simulating the passage through the stomach (T = 1’Ah)
• Approximately 1.5 hours after addition of bile and pancreatin (T = 3h)
• At the end of the digestion period (T = 6h)

Living and viable bacteria count (cfu/g) at 4 times in the GI survival model test: T = ‘Ah, T = 1’Ah, T = 3h and T =6h

Megaflora 9 evo clearly shows that after the passage of GI tract, the number of living bacteria (cfu/g) was stable with a minimum of loss. This initial decrease in the number of bacteria is better than usual expected in probiotic products.

A part of the impact which might result from the survival in the GI tract, the addition of certain ingredients to the product can damage bacteria so that, although they survive, cannot reach its optimal activity level. These cells can be counted as viable, but are not active and therefore the product loses its effectiveness.

After passing through the GI tract, the probiotic bacteria reach the small intestine, where they have to be effective at the right time and in the right place.

Metabolic or biological activity is one of the most important parameters that determines the quality of a probiotic based product. In fact, it is even more important than the amount of bacteria in the product since, from a metabolic point of view, it is useless to have a very high number of bacteria in the intestine if they are not viable.

The lactic acid production can be used as an indicator of the activity of the product since is a specific feature of probiotic activity and one of the actions that promotes health. It can be state without no doubt that the greater the production of lactic acid, the higher metabolic activity of bacteria or mixture is. Hence, measuring the production of lactic acid is considered a good method for determining activity of the probiotic strains.

The lactic acid production must be measured over time and always after a simulation of passage through the gastrointestinal tract.

When comparing the average production of lactic acid of Megaflora 9 evo with two other similar prebiotic and probiotic mixtures in concentration and gastroresistance, Megaflora 9 evo showed that after 4 hours the producing lactic acid activity was far superior.

Activity producing lactic acid of Megaflora 9 evo compared with other similar marketed probiotic mixtures with the same capacity of gastroresistance.

This result establishes that Megaflora 9 evo contains living, viable and active bacteria after GI passage.