CZ38830U1 - An individualised probiotic composition based on the analysis of microbiome and health history of an individual for humane or veterinary use - Google Patents

Description

The Industrial Property Office does not determine in the registration procedure whether the subject matter of the utility model meets the conditions for eligibility for protection pursuant to Section 1 of Act No. 478/1992 Coll.

Personalized probiotic composition based on microbiome analysis and individual health history for human or veterinary use

Technical area

The invention relates to a probiotic composition individually prepared based on microbiome analysis and taking into account the health history of the individual intended to supplement the friendly microbiota in humans or animals.

State of the art

The human gastrointestinal tract (GIT) is populated by a microbial community consisting of hundreds of different species. The intestinal microflora plays an important role not only in the digestion of food, the metabolism of endogenous and exogenous substances and the production of essential vitamins, but also in enhancing the body's immune response and preventing infections caused by pathogenic bacteria ¹ .

Human skin is home to millions of microorganisms, called the skin microbiota (Byrd, 2018) ³⁹ The skin microbiota plays an important role not only in protecting the skin from pathogenic microorganisms, but also in modulating the skin’s immune response. It is believed that agents produced by skin bacteria also contribute to healthy skin condition through positive stimulation of skin cells.

The human oral cavity is a natural environment for a diverse community of microorganisms, referred to as the oral microbiome (Dewhirst et al., 2010) ⁴⁰ . The oral microbiome is involved not only in maintaining the balance between “good” and potentially pathogenic bacteria, but also plays a key role in protecting the mucous membranes and teeth from infection, inflammation and caries. Some bacteria of the oral microbiota produce substances with antimicrobial and immunomodulatory effects, which contribute to local and general immune protection. A healthy oral microbiome is therefore considered an important factor in overall health.

The human urogenital tract, especially in women, is populated by a specific microbiota that plays a key role in protecting against pathogens and maintaining a healthy vaginal environment (Ravel et al., 2011) ⁴¹ . The dominant bacteria are mainly Lactobacillus species, which create an acidic pH and produce antimicrobial substances, thereby preventing the overgrowth of unwanted microorganisms. Disruption of this balance - for example, due to the influence of antibiotics, hormonal changes or hygiene habits - can lead to urinary tract infections, bacterial vaginosis or other complications. A stable urogenital microbiome is therefore an important component of female reproductive and urological health.

By definition, probiotics are live microbial food supplements that benefit the host by improving the balance of intestinal microflora ² . A more current definition from the perspective of human nutrition states that probiotics are live microbial food components that confer health benefits ³ . According to FAO/WHO (2002), probiotics are defined as live microorganisms which, when administered to the host in adequate amounts, confer an improvement in the host's health.

In addition to proven therapeutic properties, probiotic microorganisms must meet other requirements. Special emphasis is placed on safety ⁴ These are at least the following properties: non-pathogenic properties (including toxin production), resistance to gastric acids and bile, adhesion to intestinal epithelial cells, ability to colonize the intestine, proven beneficial effect on the host, safety. Strains must be clearly identified and at the same time deposited in an international collection of microorganisms.

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Lactic acid bacteria are important probiotic microorganisms that act not only in the human GIT, but also on the skin, in the oral cavity or urogenital tract. Representatives are classified using phenotypic characteristics, which are morphology, glucose fermentation, growth at different temperatures, lactic acid production and carbohydrate fermentation ⁵ . Probiotic lactic acid bacteria include the genera Lactobacillus, Bifidobacterium Enterococcus, Streptococcus, Leuconostoc mesenteroides, Pediococcus, Lactococcus ⁵ .

The group of lactic acid bacteria according to the newer nomenclature does not include the genus Bifidobacterium, which belongs to a completely different branch (Actinobacteria) than other lactic acid bacteria. Due to similar physiological properties, the method of use in the food industry and also with regard to the common history, it is usually assigned to this group 6.

According to current scientific studies, maintaining an appropriate content of lactobacilli and bifidobacteria not only in the intestine but also in the oral cavity is important for human health. Reducing the amount of these bacteria in the human microbiome increases the risk of metabolic disorders and other diseases 7. It has also been shown that maintaining an optimal ratio of the two basic bacterial strains Firmicutes/Bacteroidetes is important for the proper functioning of the microbiome and energy metabolism. It has been found that if an individual has a Firmicutes/Bacteroidetes ratio higher than 1 in the intestinal microbiome, they are 23% more likely to be obese 8. However, people who are very physically active (athletes) also have a higher proportion of Firmicutes. Bacteria of this strain are able to break down energy-rich compounds into simpler absorbable substrates and thus increase the energy potential of food 9. These data correspond to the data that we obtained in our research.

Both lactobacilli and bifidobacteria are present in the microbiome of a healthy person ¹⁰ . In a healthy person, bifidobacteria occur in the intestinal microbiome in the range of 2 to 14% in the digestive tract. Especially in adulthood, the amount of bifidobacteria stabilizes at 3 to 6% ^{11 ; 12} . A decrease in bifidobacteria is very often associated with a number of diseases, such as atopic dermatitis, allergies, increased risk of oncological findings, etc. A reduced amount of bifidobacteria is characteristic of patients with IBS ¹³ or in patients with allergies, in whom probiotic supplementation helps to reduce allergy symptoms ¹⁴ .

The positive effect of probiotic bacteria on human health is confirmed by numerous clinical studies and a large number of professional scientific publications.

Patent applications currently deal mainly with methods of microbiome analysis. For example, patent RU 2015146401, where the authors describe a method of microbiome culture analysis using various culture media. Document CN 109115570 concerns a method of fully automated detection of microbiome dysbiosis regardless of probiotic supplementation, EP 2985350 describes a modification of the microbiome analysis method by adding a defined amount of standard, which allows for more precise quantification of individual bacterial components in the microbiome. Document US 2017268046 concerns a method of collecting and stabilizing a stool sample and its subsequent analysis according to the content of nucleic acid sequences. In document RU 2666608, the authors describe a method of treating mycosis of the feet, where the determination of the content of lactobacilli, bifidobacteria and E.coli is part of the selection of antifungal treatment. US 2017356029 deals with microbiome typing in relation to autism, but without any connection to specific probiotic supplementation.

Some patent applications focus on screening lactobacilli and bifidobacteria with a special emphasis on their antagonistic effect against selected bacterial pathogens, such as RU 2177152 and RU 2412989. RU 2428468 is based on the selection of probiotic bacteria capable of inhibiting pathogenic bacteria isolated from the patient's intestine. Russian patent application RU 2002125639 concerns the preparation of suppositories using two bacterial strains regardless of the composition of the microbiome.

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Other documents regulate the preparation of a probiotic preparation containing bifidobacteria, but again without regard to microbiome analysis. Patent application RU 2126043 addresses the method of preserving and isolating strains from stool samples of a healthy person and their subsequent multiplication and administration to a donor during the period of illness. This is essentially the preparation of autogenous probiotics. Here too, the adjustment of the composition of the probiotic preparation depending on the composition of the microbiome of the recipient of the probiotics is not addressed. Korean patent application KR 20190025180 focuses on the analysis of the microbiome of animals with subsequent health advice, but not on the design of the composition of probiotics. Document TW 201732724 concerns the method of developing a probiotic product and its preparation system, which is not on an individual basis, but targets wider groups of patients with regard to the diagnosis and regional specificities of the microbiome.

Nowadays, all probiotics in the form of food supplements, medical devices, veterinary dietetic products, foods, foods for special purposes or cosmetics are designed on a universal principle that allows for large-scale production of such preparations. Probiotic preparations do not respect the individual needs of their users. Although there are probiotics on the market specialized for various indications, an individualized approach is not respected.

Microbiome characterization is currently performed mainly by sequencing analysis ¹⁷ , which quantifies bacterial-specific 16S rRNA gene sequences. Another commonly used procedure for determining the relative amount of bacterial DNA of selected bacterial species, genera or strains is the Real Time PCR method 8 . A conservative method for characterizing the intestinal microbiome is standard cultivation ¹⁵ , where the number of bacterial cells is determined by culturing a diluted sample on selection media, with the possibility of subsequent identification of the biochemical profile ¹⁶ .

Nowadays, all probiotics in the form of food supplements, medical devices, medicines, veterinary dietetic products, veterinary feed, food, food or cosmetics for special purposes are designed on a universal principle that allows for large-scale production of such preparations. Probiotic preparations do not respect the individual needs of their users with regard to the current state or composition of the microbiome. Although there are probiotics on the market specialized for various indications, an individualized approach is not respected.

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⁴ FRIC, P. Probiotics and prebiotics - Renaissance of a therapeutic principle. 2007. 237 to 270 ⁵ HOLZAPFEL, W. et al. Taxonomy and important features of probiotic microorganisms in food and nutrition. 2001. 365S to 373S

BORRIELLO, S.P. et al. Safety of probiotics that contain lactobacilli or bifidobacteria. Clin Infect Dis, v. 36, n. 6, p. 775 to 780, Mar 2003. ISSN 1537-6591. Available from: https://www.ncbi.nlm.nih.gov/pubmed/12627362.

CROVESY, L. et al. Effect of Lactobacillus on body weight and body fat in overweight subjects: a systematic review of randomized controlled clinical trials. Int J Obes (Lond), c.

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41, n. 11, p. 1607 to 1614, 11 2017. ISSN 1476-5497. Available in:

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⁸ KOLIADA, A. et al. Association between body mass index and Firmicutes/Bacteroidetes ratio in an adult Ukrainian population. BMC Microbiol, v. 17, n. 1, p. 120, 05 2017. ISSN 1471-2180. Available at: https://www.ncbi.nlm.nih.gov/pubmed/28532414.

⁹ MACH, N.; FUSTER-BOTELLA, D. Endurance exercise and gut microbiota: A review. J Sport Health Sci, v. 6, n. 2, p. 179 to 197, Jun 2017. ISSN 2213-2961. Available at: https://www.ncbi.nlm.nih.gov/pubmed/30356594.

¹⁰ SGHIR, A. et al. Quantification of bacterial groups within human fecal flora by oligonucleotide probe hybridization. Appl Environ Microbiol, v. 66, n. 5, p. 2263 to 2266, May 2000. ISSN 0099-2240. Available in:

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¹¹ HOPKINS, M.; SHARP, R.; T MACFARLANE, G. Age and disease-related changes in intestinal bacterial populations assessed by cell culture, 16S rRNA abundance, and community cellular fatty acid profiles. 2001. 198 to 205 ¹² SATOKARI, R. et al. Molecular Approaches for the Detection and Identification of Bifidobacteria and Lactobacilli in the Human Gastrointestinal Tract. 2003. 572 to 584 ¹³ TAVERNITI, V.; GUGLIELMETTI, S. Methodological issues in the study of intestinal microbiota in irritable bowel syndrome. 2014. 8821 to 8836 ¹⁴ AKAY, H. et al. The relationship between bifidobacteria and allergic asthma and/or allergic dermatitis: A prospective study of 0-3 year-old children in Turkey. 2014.

¹⁵ DELGADO, S.; SUÁREZ, A.; MAYO, B. Identification of dominant bacteria in feces and colonic mucosa from healthy Spanish adults by culturing and by 16S rDNA sequence analysis. Dig Dis Sci, v. 51, n. 4, p. 744 to 751, Apr 2006. ISSN 0163-2116. Available at: https://www.ncbi.nlm.nih.gov/pubmed/16614998 >.

¹⁶ TALAIEKHOZANI, A.; ALAEE, S.; MOHANADOSS, P. Guidelines for quick application of biochemical tests to identify unknown bacteria. 2015. 65 to 82 ¹⁷ DI SEGNI, A. et al. Guided Protocol for Fecal Microbial Characterization by 16S rRNA Amplicon Sequencing. J Vis Exp, n. 133, 03 2018. ISSN 1940-087X. Available at: < https://www.ncbi.nlm.nih.gov/pubmed/29608151.

¹⁸ THOMAS, DJ et al. Lactobacillus rhamnosus HN001 attenuates allergy development in a pig model. PLoS One, v. 6, n. 2, p. e16577, Feb 2011. ISSN 1932-6203. Available at: < https: //www.ncbi .nlm .nih.gov/pubmed/21386995 .

¹⁹ KALLIOMÁKI, M. et al. Probiotics in primary prevention of atopic disease: a randomized placebo-controlled trial. Lancet, v. 357, n. 9262, p. 1076 to 1079, Apr 2001. ISSN 01406736. Disponível em: < https://www.ncbi.nlm.nih.gov/pubmed/11297958 .

²⁰ TOSCANO, M. et al. Effect of. World J Gastroenterol, v. 23, n. 15, p. 2696 to 2704, Apr 2017. ISSN 2219-2840. Available at: https://www.ncbi.nlm.nih.gov/pubmed/28487606.

²¹ PAINEAU, D. et al. Effects of seven potential probiotic strains on specific immune responses in healthy adults: a double-blind, randomized, controlled trial. FEMS Immunol

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Med Microbiol, v. 53, n. 1, p. 107 to 113, Jun 2008. ISSN 0928-8244. Available at: https://www.ncbi.nlm.nih.gov/pubmed/18422632.

²² LEYER, G. et al. Probiotic Effects on Cold and Influenza-Like Symptom Incidence and Duration in Children. 2009. e172-9 ²³ FORSSTEN, S. et al. Influence of a probiotic mixture on antibiotic induced microbiota disturbances. 2014. 11878 to 11885 ²⁴ ENGELBREKTSON, A. et al. Probiotics to minimize the disruption of faecal microbiota in healthy subjects undergoing antibiotic therapy. J Med Microbiol, v. 58, n. Pt 5, p. 663 to 670, May 2009. ISSN 0022-2615. Available in:

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²⁵ OUWEHAND, AC et al. Probiotics reduce symptoms of antibiotic use in a hospital setting: a randomized dose response study. Vaccine, v. 32, n. 4, p. 458 to 463, Jan 2014. ISSN 1873-2518. Available at: https://www.ncbi.nlm.nih.gov/pubmed/24291194.

²⁶ GOMES, AC et al. The additional effects of a probiotic mix on abdominal adiposity and antioxidant status: A double-blind, randomized trial. Obesity (Silver Spring), v. 25, n. 1, p.

to 38, 01 2017. ISSN 1930-739X. Available in:

<https://www.ncbi.nlm.nih.gov/pubmed/28008750>.

²⁷ ANDREASEN, A. et al. Effects of Lactobacillus acidophilus NCFM on insulin sensitivity and the systemic inflammatory response in human subjects. 2010. 1831-8 ²⁸ RINGEL-KULKA, T. et al. Probiotic Bacteria Lactobacillus acidophilus NCFM and Bifidobacterium lactis Bi-07 Versus Placebo for the Symptoms of Bloating in Patients With Functional Bowel Disorders A Double-blind Study. 2011. 518 to 525 ²⁹ SCHMIDT, E. et al. Antigen-Presenting Cells Exposed to Lactobacillus acidophilus NCFM, Bifidobacterium bifidum BI-98, and BI-504 Reduce Regulatory T Cell Activity. 2010. 390 to 400 ³⁰ LAMIKI, P. et al. Probiotics in Diverticular Disease of the Colon: an Open Label Study. 2010. 31 to 36 ³¹ RINGEL-KULKA, T. et al. Lactobacillus acidophilus NCFM affects colonic mucosal opioid receptor expression in patients with functional abdominal pain - A randomized clinical study. 2014.

³² WICKENS, K. et al. Effects of Lactobacillus rhamnosus HN001 in early life on the cumulative prevalence of allergic disease up to 11 years. Pediatr Allergy Immunol, v. 29, n. 8, p. 808 to 814, 12 2018. ISSN 1399-3038. Available in:

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³³ CÁCERES, P. et al. Effects of Lactobacillus rhamnosus HN001 on acute respiratory infections and intestinal secretory IgA in children. 2010. 353 to 362 ³⁴ KONSTANTINOV, SR et al. S layer protein A of Lactobacillus acidophilus NCFM regulates immature dendritic cell and T cell functions 2008.

³⁵ SLYKERMAN, RF et al. Effect of Lactobacillus rhamnosus HN001 in Pregnancy on Postpartum Symptoms of Depression and Anxiety: A Randomized Double-blind Placebo

- 5 CZ 38830 U1 controlled Trial. EBioMedicine, v. 24, p. 159 to 165, Oct 2017. ISSN 2352-3964. Available at: https://www.ncbi.nlm.nih.gov/pubmed/28943228.

³⁶ WICKENS, K. et al. Early pregnancy probiotic supplementation with Lactobacillus rhamnosus HN001 may reduce the prevalence of gestational diabetes mellitus: a randomized controlled trial. 2017. 1 to 10 ³⁷ JANG, SE et al. Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus La-14 Attenuate Gardnerella vaginalis-Infected Bacterial Vaginosis in Mice. Nutrients, v. 9, n. 6, May 2017. ISSN 2072-6643. Available in:

<https://www.ncbi.nlm.nih.gov/pubmed/28545241.

³⁸ BO CORREA, N. et al. Treatment of Acute Diarrhea With Saccharomyces Boulardii in Infants. 2011. 497 to 501 ³⁹ Byrd, A.; Belkaid, Y.; Segre, J. The human skin microbiome. Nat. Roar. Microbiol. 2018, 16, 143 to 155.

⁴⁰ Dewhirst, FE, Chen, T., Izard, J., Paster, BJ, Tanner, ACR, Yu, W.-H., Lakshmanan, A., & Wade, WG (2010). The human oral microbiome. Journal of Bacteriology, 192(19), 5002 to 5017.

⁴¹ Ravel, J., Gajer, P., Abdo, Z., Schneider, GM, Koenig, SSK, McCulle, SL, ... & Forney, LJ (2011). Vaginal microbiome of reproductive-age women. Proceedings of the National Academy of Sciences, 108(Supplement 1), 4680-4687.

The essence of the technical solution

The deficiencies of the prior art, especially in the area of individualized production of probiotic preparations, are addressed by the probiotic composition according to the technical solution. The method of preparing this composition is based on a combination of two factors, namely the microbiome analysis of an individual's sample and his/her health history, on the basis of which a specific formulation of the component of selected probiotic cultures with a suitable excipient selected from the group of glucooligosaccharides and/or xylooligosaccharides is prepared into the resulting probiotic composition.

An individualized probiotic composition for human or veterinary use according to the technical solution is prepared based on the analysis of a sample of the intestinal, or oral, skin or urogenital microbiome of an individual. The analysis is aimed at quantitatively determining the percentage of the relative representation of bacteria of the genera Lactobacillus, Bifidobacterium in the analyzed sample. Microbiome analysis for the purposes of the technical solution is performed by culturing the sample with the possibility of subsequent identification of the biochemical profile and/or Real Time PCR and/or sequencing of a stool sample or, optionally, any combination of the above methods. The representation of other bacterial genera, species or entire taxa can be advantageously determined. At the same time, the individual's health history is obtained, i.e. a set of diseases, or. all health disorders.

According to the results of the analysis of the individual's microbiome sample, especially with regard to the relative representation of the genus Lactobacillus and preferably also Bifidobacterium, a formulation of the component of individualized probiotic cultures for the probiotic composition is prepared. The procedure is such that the individually prepared probiotic composition preferentially contains a sufficient amount of the genera Lactobacillus or Bifidobacterium bacteria, which, according to the results of the microbiome analysis, are reduced or absent in comparison with the representation of bacterial cultures in the microbiome of a healthy person. To supplement the amount of missing genera of bacteria, those probiotic bacterial strains are then preferentially selected that are clinically tested for indications obtained from the anamnesis

- 6 CZ 38830 U1 with proven health benefits for at least one of the indications from the individual's health history according to at least one piece of information from literature, internet sources and/or the result of a clinical study.

This is a unique combination - supplementation of bacteria from the group of microorganisms of the genus Lactobacillus and Bifidobacterium, the amount of which is reduced or completely absent in the patient's microbiome, in a certain percentage, according to the determined percentage of the relative representation of these bacteria in the analytical sample with preferential selection of strains clinically tested for the selected anamnesis. Finally, an auxiliary component selected from the group of glucooligosaccharides and/or xylooligosaccharides is added to the prepared component of individualized probiotic cultures for the probiotic composition.

The subject of the technical solution is an individualized probiotic composition for human or veterinary use containing a component of probiotic cultures of the genus Lactobacillus and Bifidobacterium and an auxiliary component containing soluble fiber based on glucooligosaccharides and/or xylooligosaccharides. Probiotic cultures of the genus Lactobacillus and Bifidobacterium constitute at least 80% by volume of the total volume of the probiotic cultures component.

The probiotic culture component may also contain probiotic cultures selected from the group of Streptococcus, Bacillus, Sacharomyces, Enterococcus, Lactococcus, Escherichia, Akkermansia, Faecalibacterium, Eubacterium, Leuconostoc, Pediococcus, Weizmania or mixtures thereof.

The probiotic culture component contains probiotic cultures in vital form, preferably lyophilized, in an amount of 1 x 10 ³ to 1 x 10 ¹⁴ CFU (colony forming units) per 1 g of total weight of the probiotic composition, preferably 1 x 10 9 CFU per 1 g to 200 x 10 9 CFU per 1 g of total weight of the probiotic composition. The lyophilized probiotic culture component can first be suspended in oil, especially in the case of cosmetic preparations, then subsequently mixed into the final cosmetic formulation. The oil constitutes at least 10% by weight of the total weight of the probiotic culture component. It is preferably selected from the group consisting of rapeseed oil, olive oil, sunflower oil, avocado oil, coconut oil, linseed oil, castor oil, evening primrose oil, almond oil, argan oil, grape seed oil, apricot oil, jojoba oil, caraway oil, rose oil, raspberry oil, coffee oil, plum oil, MCT oil (medium chain fatty acid oil). This suspension is highly stable when subsequently mixed into a cream, ointment, body oil, mouthwash, soap or shampoo and is capable of maintaining the viability of probiotic bacteria during use of the preparation.

The auxiliary component of the probiotic composition may advantageously also contain inulin, dextrin, maltodextrin, corn starch, potato starch, fructooligosaccharides, galactooligosaccharides, isomaltooligosaccharides, pectins, guar gum, xanthan gum, carrageenan, alginate, fruit extracts, vegetable extracts, grain extracts, legume extracts, lyophilized fruit, lyophilized vegetables, vitamins, minerals, plant and herbal extracts or mixtures thereof and other physiologically beneficial substances or substances with prebiotic potential in a maximum amount of up to 40.00% by weight.

The individualized probiotic composition according to a preferred embodiment o contains 0 to 34% by volume of probiotic cultures of the genus Bifidobacterium and 66 to 99% by volume of probiotic cultures of the genus Lactobacillus of the total volume of these cultures in the probiotic culture component, if the percentage ratio of the relative representation of lactobacilli and bifidobacteria in the analyzed microbiome sample is lower than 0.099.

If the percentage ratio of the relative representation of lactobacilli and bifidobacteria in the sample of the analyzed microbiome of an individual is in the range of 0.1 to 0.799, the probiotic composition preferably contains 2 to 49% probiotic cultures of the genus Bifidobacterium and 51 to 98% probiotic cultures of the genus Lactobacillus, preferably 35% bifidobacteria and 65% lactobacilli of the total volume of these cultures in the probiotic cultures component.

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If the percentage ratio of the relative representation of lactobacilli and bifidobacteria in the sample of the analyzed microbiome of an individual is between 0.8 and 1.099, the probiotic composition contains 20 to 80% probiotic cultures of the genus Bifidobacterium and 20 to 80% probiotic cultures of the genus Lactobacillus, preferably 50% bifidobacteria and 50% lactobacilli of the total volume of these cultures in the probiotic cultures component.

If the percentage ratio of the relative representation of lactobacilli and bifidobacteria in the sample of the analyzed microbiome of an individual is higher than 1.1, the probiotic composition contains 20 to 99% probiotic cultures of the genus Bifidobacterium and 1 to 80% probiotic cultures of the genus Lactobacillus, preferably 75% bifidobacteria and 25% lactobacilli of the total volume of these cultures in the probiotic cultures component.

The subject of the technical solution is also a food, pharmaceutical or veterinary product containing an individualized probiotic composition according to the technical solution, for example in the form of a food, confectionery, food supplement, product for special nutrition, food, beverage, e.g. yogurt, kefir, it can be advantageously in the form of a tablet, capsule, powder, gel, suppository, solution, medical device, hygiene product or veterinary feed and veterinary dietetic preparation, for example in the form of granules or granulate.

The subject of the technical solution is also a cosmetic product containing a probiotic composition according to the technical solution, for example in the form of a cream, ointment, mouthwash, cosmetic oil, soap, shampoo, cosmetic suspension or emulsion.

Examples of implementing a technical solution

Example 1

Capsules containing an individualized probiotic composition

The ratio of lactobacilli and bifidobacteria in the individual's stool sample was determined by sequencing molecular biological analysis and subsequently by Real Time PCR to a value of 0.084. Based on this ratio, it was decided that 25% of the volume of the probiotic cultures corresponding to the daily dose of probiotics (35 x 109 CFU) would be bifidobacteria and 75% of the volume of lactobacilli. The client, 45 years old, female, had a history of allergies and dermatitis. Therefore, probiotic bacteria that have clinical studies or other published results demonstrating a therapeutic effect with respect to allergies ¹⁸ and dermatitis ¹⁹ were preferentially used, supplemented with other probiotic species to increase the overall bacterial diversity in the intestine. Fructooligosaccharide from chicory - inulin was used as an auxiliary prebiotic substance. The mixture of commercially available probiotic cultures according to Table 1 was homogenized in a dry lyophilized state in a laboratory homogenizer for 10 min. Filling into capsules was carried out using a manual capsule filler in a laboratory with controlled air humidity of up to 35% and temperature of up to 22 °C.

The resulting probiotic composition with a capsule filling weight of 450 mg, representing a daily dose of the food supplement, was filled into enteric-coated size 0 capsules using a manual encapsulator. The resulting composition of the individualized probiotic composition was as follows:

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Table 1

Type of probiotic bacteria Trade name Manufacturer Stock bacteria concentration CFU/g Vol. % of bacteria in composition i (capsules) Weight of individual components in the composition (mg) Lactobacillus rhamnosus (tested for allergy suppression) HN001 Danisco 4.50 x 10 ¹¹ 30.62 2.33 Lactobacillus acidophilus (clinical trials for immune-allergy regulation) NCFM Danisco 2.00 x 10 ¹¹ 15.31 2.63 Lactobacillus reuterii (clinical trials for immune regulation- allergies) BioCareCopenhagen 3.00 x 10 ¹¹ 8.66 0.88 Lactobacillus acidophilus (supports faster immune response through IgG antibodies) LA-14 Danisco 2.00 x 10 ¹¹ 15.81 2.63 Lactobacillus plantarum LP-ONLLI Novanate 4.00 x 10 ¹¹ 2.30 0.20 Lactobacillus acidophilus Nowanat 1.50 x 10 ¹¹ 2.30 0.53 Streptococcus thermophilus ST37 Synbiotech 1.00Ex 10 ¹¹ 2.00 1.05 Bifidobacterium breve BB8 Novanate 3.00 x 10 ¹¹ 4.90 5.83 Bifidobacterium bifidum BB47 Novanate 2.00 x 10 ¹¹ 11.00 19.69 Bifidobacterium lactis, Bifidobacterium bifidum 1:1 mixture Bb-02 Danisco 3.40 x 10 ¹¹ 6.16 6.43 Bifidobacterium infantis BI17 Synbiotech 1.00 x 10 ¹¹ 1.72 4.38 Bifidobacterium animalis BAL06 Synbiotech 1.00 x 10 ¹¹ 1.22 4.38 Filler Glucooligosaccharides (Soluble Insoluble Fiber) Promiser Azelis 399.07

Evaluation: After 5 months of use, the skin problems disappeared. At the same time, 5 allergic reactions to dust characterized by swollen mucous membranes subsided. PCR analysis revealed that the ratio of Lactobacilli and Bifidobacteria in the sample increased to 0.75.

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Example 2

Tablets containing an individualized probiotic composition

The ratio of lactobacilli and bifidobacteria in the stool sample was determined by sequencing molecular biological analysis to be 0.93 and the number of bacterial species in the sample was also determined. The proportion of probiotic bacteria in the resulting probiotic composition was determined to be 50% by volume of bifidobacteria, a component of probiotic cultures corresponding to the daily dose, and 50% by volume of lactobacilli. A comprehensive molecular biological analysis showed a significantly reduced diversity of the intestinal microbiome of the tested person in the number of 248 detectable bacterial species. The reason could be the recent intensive antibiotic therapy against H. pylori, which was mentioned in the anamnesis. Furthermore, the client stated that he suffers from frequent diarrhea. For these reasons, the daily dose was determined to be 45 x 10 ⁹ CFU, fortified mainly with bacterial species that are able to reduce the negative effect of antibiotics on the bacterial microbiota and reduce diarrhea ^{23; 24; 25} . Due to diarrheal conditions, a mixture of gluco-oligosaccharides and xylo-oligosaccharides in a ratio of 1:1 was used as an auxiliary prebiotic substance in the amount of 10% of the total capsule content and SiO2 in the amount of 1% of the total tablet content, which supports the adhesion of probiotic bacteria to the intestinal epithelium. The mixture was supplemented with 0.5% of the total tablet content of magnesium stearate and also with banana lyophilisate.

The composition of the resulting composition corresponded to the data in Table 2. The individualized probiotic composition weighing 450 mg adjusted into oval tablets corresponded to the daily dose of the food supplement.

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Table 2

Type of probiotic bacteria Trade name Manufacturer Stock bacteria concentration CFU/g Vol.% of bacteria in the composition (tablet) Weight of individual components in the composition (mg) Bifidobacterium lactis (reduction of antibiotic-induced diarrhea) Bi-07 Danisco 4.00 x 10 ¹¹ 12.75 8.40 Bifidobacterium lactis (reduction of antibiotic-induced diarrhea) BL04 Danisco 3.00 x 10 ¹¹ 12.75 11.30 Bifidobacterium bifidum BB47 Novanate 2.00 x 10 ¹¹ 5.25 5.60 Bifidobacterium lactis B420 Danisco 4.00 x 10 ¹¹ 10.25 2.80 Bifidobacterium animalis BAL06 Synbiotech 1.00 x 10 ¹¹ 4.75 11.30 Bifidobacterium breve BB8 Novanate 3.00 x 10 ¹¹ 4.25 3.80 Lactobacillus acidophilus NCFM Danisco 2.00 x 10 ¹¹ 24.30 22.50 Lactobacillus sporogenes Lactospor e Sabinsa 1.50 x 10 ¹⁰ 2.40 30.00 Lactobacillus paracasei LPC48 Synbiotech 1.00 x 10 ¹¹ 4.00 11.30 Streptococcus thermophilus ST37 Synbiotech 1.00 x 10 ¹¹ 10.00 22.50 Lactococcus lactis subsp. Lactis LLL970 Synbiotech 1.00 x 10 ¹¹ 10.00 11.30 Lactobacillus casei LC18 Novanate 1.00 x 10 ¹¹ 6.07 11.30 Lactobacillus cripatus Novanate 5.00 x 10 ¹⁰ 3.25 13.50 Filler A mixture of gluco-oligosaccharides and xylo-oligosaccharides in a ratio of 1:1 45.00 SiO ₂ 4.50 Magnesium stearate 2.25 Banana freeze-dried 232.65

Evaluation after 3 months: Increased microbiome diversity to 326 detectable 5 bacterial species in the sample. Diarrhea completely resolved.

Example 3 individualized probiotic composition in powder form

The ratio of lactobacilli to bifidobacteria in the stool sample was determined by molecular biological analysis to be 0.237. Based on this ratio, it was decided that 35% of the volume of probiotic cultures corresponding to the daily dose of probiotics (35 x 10 ⁹ CFU) would be

-11 CZ 38830 UI bifidobacteria and 65% vol. lactobacilli. The total number of bacterial species in the sample and the level of cholesterol in the blood were also determined. The client, 39 years old, male, had a history of persistent constipation, bloating, and flatulence. Therefore, probiotic bacteria were preferentially used that have clinical studies or other published results demonstrating a therapeutic effect against constipation, supplemented with another 5 probiotic species to reduce digestive discomfort ²⁸ and increase the overall bacterial diversity in the intestine ²¹ . Inulin, which has an anti-constipating effect, was used in an amount of 10% of the total capsule content, and was supplemented with an auxiliary prebiotic substance - xylooligosaccharides.

The resulting probiotic composition weighing 500 mg, corresponding to a daily dose of a food supplement, intended for the production of a solution, or for dissolution in a lukewarm drink, had the composition shown in Table 3.

Table 3

Type of probiotic bacteria Trade name Manufacturer Stock bacteria concentration CFU/g Vol.% of bacteria in the composition (powder) Weight of individual ingredients in the composition (mg) Lactobacillus acidophilus (reduces the transit time of food through the digestive tract) LA-14 Danisco 2.00 x 10 ¹¹ 39.00 34.10 Lactobacillus acidophilus (reduces bloating) NCFM Danisco 2.00 x 10 ¹¹ 9.10 8.00 Lactobacillus paracasei LPC48 Synbiotech 1.00 x 10 ¹¹ 9.10 15.90 Lactobacillus plantarum LP-ONLLI Novanate 2.00 x 10 ¹¹ 2.60 2.30 Lactobacillus bulgaricus Novanate 5.00 x 10 ¹¹ 2.60 9.10 Lactobacillus cripatus Novanate 5.00 x 10 ¹¹ 2.60 9.10 Bifidobacterium lactis (reduces flatulence) Bi-07 Danisco 4.00 x 10 ¹¹ 10.50 4.60 Bifidobacterium lactis, Bifidobacterium bifidum 1:1 mixture Bb-02 Danisco 4.50 x 10 ¹¹ 10.50 4.10 Bifidobacterium breve BB8 Novanate 3.00 x 10 ¹¹ 7.00 4.10 Bifidobacterium bifidum BB47 Novanate 2.00 x 10 ¹¹ 3.50 3.10 Bifidobacterium infantis BI211 Novanate 3.00 x 10 ¹¹ 3.50 2.00 Filler Inulin 50.00 Xylooligosaccharides 353.60

Evaluation of the client's condition: After 2 months of use, bowel movements improved to 1x in two days, the debilitating constipation subsided. At the same time, the ratio of Lactobacilli/Bifidobacteria in the sample increased to 1.1.

- 12 CZ 38830 UI

Example 4

Ointment containing an individualized probiotic composition

The content of the Staphylococcus aureus strain in a skin swab sample from the facial area was determined by molecular biological analysis to be 1.8% of the total bacterial DNA content and the content of the bacterial genus Cutihacterium was determined to be 26%. The ratio of the genus Lactobacillus and Bifidobacterium in the skin microbiome was determined to be 2.9. In addition, the person showed increased formation of wrinkles and red spots on the face. Based on this ratio, it was decided that 70% of the daily dose of bifidobacteria and lactobacilli in the probiotic mixture by weight would be bifidobacteria and 30% by volume lactobacilli. Taking into account the anamnesis, a component of individualized probiotic cultures for a cosmetic product was prepared. The probiotic mixture was prepared according to the composition in Table 4. A mixture of polyphenols from olives, red wine, pomegranate, orange and green tea was used as an auxiliary prebiotic component.

The resulting component of individualized lyophilized probiotic cultures with the composition according to Table No. 4 by weight was homogenized with 1.5 ml of macadamia oil. This oil suspension was homogenized into an ointment with a volume of 50 ml with the composition according to Table 5.

Table 4

Type of probiotic bacteria Trade name Manufacturer Stock bacteria concentration CFU/g Weight of individual ingredients (mg) Bifidobacterium lactis Ll-23 Danisco 1.00 X 10 ¹¹ 35.00 Bifidobacterium animalis HN019 Dancing 1.00 X 10 ¹¹ 35.00 Lactobacillus bulgaricus Lb-87 Danisco 1.00 X 10 ¹¹ 20.00 Lactobacillus casei LC11 Danisco 1.00 X 10 ¹¹ 10.00 mixture of polyphenols Oxyynea Phytexia 50.00 Glucooligosaccharides Promiser Azelis 100.00

Table 5

Folder Quantity % wt. Water 97.40 PEG-14 dimethicone 1.00 Carbomer 0.80 Sodium hydroxymethylglycinate 0.60 Hyaluronic acid 0.20

Evaluation: After monthly application, there was a noticeable reduction in the number and size of reddened lesions on the skin. PCR showed a decrease in Staphylococcus aureus in the sample to 0.3%.

Example 5

Veterinary dietetic product containing an individualized probiotic composition in the form of granules

The ratio of lactobacilli to bifidobacteria in a stool sample from a dog (Australian Shepherd breed) was determined by molecular biological sequencing analysis to be 0.025. Based on this ratio,

-13 CZ 38830 UI it was decided that 25% vol. of the probiotic cultures corresponding to the daily dose of probiotics will consist of bifidobacteria and 75% vol. of lactobacilli. The total number of bacterial species in the sample was also determined. A comprehensive molecular biological analysis showed a reduced diversity of the intestinal microbiome. In the anamnesis, the dog owner reported frequent infections and significantly reduced immunity with three 5 infections in the last 6 months. Due to the reduced diversity, the daily dose of probiotic bacteria was set at 45xlO ⁹ CFU. Probiotic bacteria that have clinical studies or other published results demonstrating a therapeutic effect with respect to respiratory tract diseases ^{22; 32; 33; 34} and have a positive effect on strengthening mucosal immunity, when the effect is expected in animals as well, were preferentially used in the probiotic composition. Due to reduced immunity, 100 mg of beta-glucan from oyster mushroom was added to the composition. Glucooligosaccharide fiber was used as an auxiliary prebiotic substance in an amount of 10% of the total content of the mixture, which supports the adhesion of probiotic bacteria to the intestinal epithelium, supplemented with carrot lyophilisate.

The resulting probiotic composition weighing 950 mg, prepared into granulate form by dry granulation, had the composition according to Table 6.

Table 6

Type of probiotic bacteria Trade name Manufacturer Stock bacteria concentration CFU/g Vol.% of bacteria in the composition (granulate) Weight of individual ingredients in the composition (mg) Lactobacillus acidophilus (supports faster immune response through IgG antibodies) LA-14 Danisco 2.00 x 10 ¹¹ 31.67 33.80 Lactobacillus rhamnosus (increased secretion of IgA antibodies) HN001 Danisco 4.50 x 10 ¹¹ 15.83 7.50 Lactobacillus acidophilus (reduction of duration of respiratory illness, regulation of dendritic cell maturation and T cell function) NCFM Danisco 2.00 x 10 ¹¹ 8.80 8.40 Lactobacillus plantarum LPONLLI Novanate 2.00 x 10 ¹¹ 8.90 8.40 Streptococcus thermophilus ST37 Synbiotech 1.00 x 10 ¹¹ 5.00 16.90 Lactobacillus cripatus Novanate 5.00 x 10 ¹⁰ 9.90 33.80 Bifidobacterium lactis B420 Danisco 4.00 x 10 ¹¹ 21.60 12.70 Bifidobacterium lactis, Bifidobacterium bifidum 1:1 mixture Bb-02 Danisco 4.50 x 10 ¹¹ 1.95 0.50 Bifidobacterium bifidum BB47 Novanate 2.00 x 10 ¹¹ 1.45 1.70 Filler 123.70 xylooligosaccharides 40.00 Betaglucan 100.00 Glucooligosaccharides 545.00 Yeast protein 141.30

-14 CZ 38830 U1

Evaluation: Half a year of supplementation with the feed resulted in the dog having only one respiratory episode in the six-month period monitored. PCR analysis revealed that the ratio of Lactobacilli to Bifidobacteria in the sample increased to 0.38.

Example 6

Gelatin capsules containing an individualized probiotic composition

The ratio of lactobacilli and bifidobacteria in the urine sample was determined by Real Time PCR analysis to be 0.88. The total number of bacterial species in the sample was also determined. The proportion of probiotic bacteria in the resulting composition was determined to be 32% by volume of bifidobacteria as a component of probiotic cultures corresponding to the daily dose and 68% by volume of lactobacilli in the daily dose of probiotic bacteria in a capsule as a food supplement. The client was a woman in the 5th month of pregnancy. For this reason, the daily dose was determined to be 25 x 109 CFU. The probiotic composition preferentially used probiotic bacteria that have clinical studies or other published results demonstrating a positive effect on the developing immune system of the fetus ³² or on its overall development and overall well-being of the expectant mother ³⁵ including support for the prevention of gestational diabetes ³⁶ . Furthermore, when selecting probiotic bacteria, emphasis was placed on digestive comfort ²⁰ , supporting the client's immune system, and also on supporting a healthy vaginal microbiota ³⁷ . Soluble corn fiber - glucooligosaccharides - was used as a prebiotic excipient.

The resulting individualized probiotic composition weighing 450 mg, adjusted into size 0 gelatin capsules, corresponding to the daily dose of the food supplement, had the composition according to Table 7.

- 15 CZ 38830 UI

Table 7:

Type of probiotic bacteria Trade name Manufacturer Stock bacteria concentration CFU/g Percentage of bacteria in a tobole Weight of individual ingredients in daily dose (mg) Lactobacillus rhamnosus (reducing the prevalence of allergies in the child, preventing gestational diabetes, reducing the incidence of postpartum depression) HN001 Danisco 4.50 x 10 ¹¹ 28.71 7.60 Lactobacillus acidophilus NCFM Danisco 2.00 x 10 ¹¹ 14.75 8.50 Lactobacillus acidophilus (to relieve symptoms of bacterial vaginosis) LA-14 Danisco 2.00 x 10 ¹¹ 17.35 8.50 Streptococcus thermophilus ST37 Synbiotech 1.00 x 10 ¹¹ 5.00 8.50 Lactobacillus plantarum LP-ONLLI Novanate 2.00 x 10 ¹¹ 7.20 4.30 Bifidobacterium lactis B420 Danisco 4.00 x 10 ¹¹ 13.10 4.00 Bifidobacterium bifidum BB47 Novanate 2.00 x 10 ¹¹ 6.18 4.00 Bifidobacterium lactis Bi-07 Danisco 4.00 x 10 ¹¹ 5.08 2.00 Bifidobacterium lactis, Bifidobacterium bifidum 1:1 mixture Bb-02 Danisco 4.50 x 10 ¹¹ 4.06 0.90 Bifidobacterium breve BB8 Novanate 3.00 x 10 ¹¹ 3.04 1.30 Filler Glucooligosaccharides (soluble corn fiber) Promiser Azelis 400.40

Conclusion: There was an increase in the ratio of lactobacilli to bifidobacteria in the sample to 1.04.

Example 7

Veterinary dietetic product containing an individualized probiotic composition in powder form

The ratio of lactobacilli and bifidobacteria in a stool sample from a golden retriever dog was determined by molecular biological sequencing analysis to be 0.035. Based on this ratio, it was decided that 25% of the volume of the probiotic cultures corresponding to the daily dose of probiotics would be bifidobacteria and 75% of the volume of lactobacilli. The total number of bacterial species in the sample was also determined. A comprehensive molecular biological analysis showed a reduced diversity of the animal's intestinal microbiome, namely 205 detectable bacterial species. The dog's owner reported frequent respiratory tract infections in the animal's history and, according to the veterinarian's examination, also reduced immunity of the animal. Due to the reduced diversity, the daily dose of probiotic bacteria was set at 15 x 10 ⁹ CFU. Probiotic bacteria that have clinical studies or other published results demonstrating

-16 CZ 38830 UI therapeutic effect with respect to respiratory tract diseases ^{22; 32; 33; 34} and have a positive effect on strengthening mucosal immunity regardless of whether these are studies on humans or animals. Due to reduced immunity, 100 mg of beta-glucan from oats and 40 mg of granulated dried bovine colostrum were added to the composition. As an auxiliary prebiotic substance, 5 soluble corn fiber based on glucooligosaccharides was used in an amount of 10% of the total capsule content, supplemented with carrot lyophilisate and meat flavor.

The resulting individualized probiotic composition weighing 450 mg, prepared into a powder form corresponding to the daily dose, had the composition according to Table 8.

Table 8

Type of probiotic bacteria Trade name Manufacturer Stock bacteria concentration CFU/g Percentage of bacteria in the daily dose Weight of individual ingredients in daily dose (mg) Lactobacillus acidophilus (supports faster immune response through IgG antibodies) LA-14 Danisco 2.00 x 10 ¹¹ 31.63 33.80 Lactobacillus rhamnosus (increased secretion of IgA antibodies) HN001 Danisco 4.50 x 10 ¹¹ 15.83 7.50 Lactobacillus acidophilus (regulation of dendritic cell maturation and T cell function) NCFM Danisco 2.00 x 10 ¹¹ 10.92 8.40 Lactobacillus plantarum LPONLLI Novanate 2.00 x 10 ¹¹ 8.70 8.40 Streptococcus thermophilus ST37 Synbiotech 1.00 x 10 ¹¹ 5 16.90 Lactobacillus cripatus Novanate 5.00 x 10 ¹⁰ 7.92 33.80 Bifidobacterium lactis B420 Danisco 4.00 x 10 ¹¹ 21.37 12.70 Bifidobacterium lactis, Bifidobacterium bifidum 1:1 mixture Bb-02 Danisco 4.50 x 10 ¹¹ 1.17 0.50 Bifidobacterium bifidum BB47 Novanate 2.00 x 10 ¹¹ 2.46 1.70 Filler Colostrum bovinni granulated 40.00 Betaglucan 100.00 Glucooligosaccharides 45.00 Carrot freeze-dried Promiser Azelis 140.00 Meat flavor Aro 1.4.00

Conclusion: 3 months of supplementation led to an increase in the number of detectable bacterial species from 15 in the stool sample to 312.

-17 CZ 38830 U1

Example 8

Veterinary dietetic product containing an individualized probiotic composition in the form of a powder mixture

The ratio of lactobacilli and bifidobacteria in a domestic cat stool sample was determined by molecular biological analysis using Real Time PCR to be 0.016. The total number of bacterial species in the sample was also determined. The proportion of probiotic bacteria in the resulting probiotic composition was designed to be 25% by volume of the probiotic culture component, corresponding to a daily dose of 10 bifidobacteria and 75% by volume of lactobacilli. Due to the low weight of the animal, the daily dose of probiotics was set at 2 x 109 CFU. The cat had a history of increased frequency of diarrhea. Therefore, S. boulardii ³⁸ was used in the probiotic composition. Furthermore, probiotic species of bacteria aimed at increasing bacterial diversity in the intestine with an emphasis on optimizing digestion and strengthening overall immunity were used in the probiotic composition ^{21; 22; 34} . As an auxiliary prebiotic substance, 15 fructooligosaccharide from chicory - inulin and soluble corn fiber were used.

The resulting probiotic composition of the powder mixture weighing 1000 mg, corresponding to the daily dose, filled into disposable sachets, had the composition according to Table 10A.

- 18 CZ 38830 UI

Table 9:

Type of probiotic bacteria Trade name Manufacturer Stock bacteria concentration CFU/g Percentage of bacteria in the daily dose (1 sachet) Weight of individual ingredients in daily dose (mg) Lactobacillus acidophilus (supports faster immune response through IgG antibodies) LA-14 Danisco 2x 10 ¹¹ 18.75 14.20 Lactobacillus acidophilus (regulation of dendritic cell maturation and T cell function) NCFM Danisco 2x 10 ¹¹ 15.00 15.00 Lactobacillus rhamnosus HN001 Danisco 4.5 x 10 ¹¹ 15.00 6.70 Lactobacillus plantarum LP-ONLLI Novanate 4x 10 ¹¹ 11.50 3.80 Streptococcus thermophilus ST37 Synbiotech 1 x 10 ¹¹ 10.00 15.00 Lactococcus lactis subsp. Lactis LLL970 Synbiotech 1 x 10 ¹¹ 2.50 7.50 Lactobacillus casei LC122 Synbiotech 1 x 10 ¹¹ 8.75 7.50 Lactobacillus bulgaricus Novanate 5 x 1O ¹⁰ 3.50 15.00 Bifidobacterium lactis, Bifidobacterium bifidum 1:1 mixture Bb-o2 Danisco 4.5 x 10 ¹¹ 5.50 2.80 Bifidobacterium lactis Bi-07 Danisco 4.00 x 10 ¹¹ 6.25 1.60 Bifidobacterium bifidum BB47 Novanate 2x 10 ¹¹ 10.00 10.10 Bifidobacterium breve BB8 Novanate 3 x 10 ¹¹ 3.25 0.80 Saccharomyces boulardii (anti-diarrheal effects) S. boulardii Lallemand 1 x 10 ¹¹ 2.50 2.50 Filler 102.5 Inulin Orafti GL Azelis 390.9 Corn Soluble Fiber Promiser Azelis 500 Meat flavor Aroma of meat Aro 2.00

Evaluation: After two months of use, diarrhea completely disappeared. The ratio of lactobacilli 5 and bifidobacteria in the sample increased to 0.29.

-19 CZ 38830 UI

Example 9

Mouthwash containing an individualized probiotic composition

The content of the Streptococcus mutans strain in the saliva sample was determined by molecular biological analysis to be 0.2% of the total bacterial DNA content and the content of the bacterial genus Fusobacterium nucleatum was determined to be 26%. The ratio of lactobacilli and bifidobacteria in the saliva microbiome was determined to be 0.9. In addition, the person showed increased dental caries and periodontitis. Based on this result and the medical history, a component of individualized probiotic cultures for a cosmetic composition in the form of a mouthwash was prepared. The probiotic mixture was prepared according to the composition in Table No. 10 and was dissolved in almond oil with a volume of 9 ml. This mixture was filled into a cap with a punctured bottom, which is screwed to the mouthwash bottle. The composition of the mouthwash is shown in Table No. 11. Before using the mouthwash, the bottom of the cap is punctured and the suspension of probiotic cultures is poured into the mouthwash. Xylooligosaccharides were used as an auxiliary prebiotic component. Based on this ratio, it was decided that 30% by weight of the daily dose of the probiotic component of the composition would consist of bifidobacteria and 70% of lactobacilli.

The resulting component of individualized lyophilized probiotic cultures according to Table No. 10 by weight was homogenized with 9 ml of almond oil. This oil suspension was homogenized into a mouthwash in a volume of 100 ml with the composition according to Table 11.

Table 10

Type of probiotic bacteria Trade name Manufacturer Stock bacteria concentration CFU/g Weight of individual ingredients (mg) Streptococcus salivarius SAL21 Fermatism 1.00 X 10 ¹¹ 50.00 Bifidobacterium animalis HN019 Dancing 1.00 X 10 ¹¹ 30.00 Lactobacillus salivarius LS33 LS33 Danisco 1.00 X 10 ¹¹ 50.00 Lactobacillus casei LC11 Danisco 1.00 X 10 ¹¹ 20.00 Xylooligosaccharides xos Porta 100.00 Glucooligosaccharides Promiser Azelis 50.00

Table 11

Ingredients (INCI) in the bottle g/100g Water 67.6 Glycerin 22.00 Polyglycerin-3 8.00 Polysorbate 80 1.50 Aloe barbadensis leaf juice 0.25 Menthol 0.20 Aroma 0.20 Sodium sorbate 0.15 Xylooligosaccharides 0.10 Total in the bottle 100.00

-20 CZ 38830 U1

Evaluation: After 3 months of using the mouthwash once a day before bed, a reduction in the abundance of Fusobacterium nucleatum in the sample to 0.15% was found. Streptococcus mutans was reduced to 0.14% in the sample.

Industrial applicability

The individualized probiotic composition for human or veterinary use according to the technical solution takes into account the current individual representation of probiotic bacteria 10 in the microbiota of an individual and appropriately supplements, in particular, bacteria from the group of microorganisms of the genus Lactobacillus and Bifidobacterium in the microbiota of an individual to the physiological level of a healthy person. Thanks to the content of preferential selection of strains clinically tested for a certain anamnesis of the individual, it contributes to maintaining his healthy microbiota and also improving his health status.

Claims (13)

PROTECTION CLAIMS 1. An individualized probiotic composition for human or veterinary use, characterized in that it contains a probiotic culture component containing the genus Lactobacillus and Bifidobacterium in vital form in an amount of 1 x 10 ³ to 1 x 10 ¹⁴ CFU per 1 g of total weight of the probiotic composition, wherein the probiotic cultures of the genus Lactobacillus and Bifidobacterium constitute at least 80% by volume of the total volume of the probiotic culture component and an auxiliary component containing soluble fiber based on glucooligosaccharides and/or xylooligosaccharides. 2. Individualized probiotic composition according to claim 1, characterized in that the probiotic culture component also contains probiotic cultures selected from the group of Streptococcus, Bacillus, Sacharomyces, Enterococcus, Lactococcus, Escherichia, Akkermansia, Faecalibacterium, Eubacterium, Leuconostoc, Pediococcus, Weizmania or mixtures thereof. 3. The individualized probiotic composition according to claims 1 to 2, characterized in that the probiotic culture component is suspended in oil, where the oil constitutes at least 10% by weight of the total weight of the probiotic culture component. 4. The individualized probiotic composition according to claim 3, characterized in that the oil is selected from the group consisting of rapeseed oil, olive oil, sunflower oil, avocado oil, coconut oil, linseed oil, castor oil, evening primrose oil, almond oil, argan oil, grape seed oil, apricot oil, jojoba oil, caraway oil, rose oil, raspberry oil, coffee oil, plum oil, and oil containing medium-chain fatty acids. 5. Individualized probiotic composition according to claim 1 to 4, characterized in that the auxiliary component contains inulin, dextrin, maltodextrin, corn starch, potato starch, fructooligosaccharides, isomaltooligosaccharides, galactooligosaccharides, pectins, beta-glucans, guar gum, xanthan gum, carrageenan, alginate, fruit extracts, vegetable extracts, grain extracts, legume extracts, freeze-dried fruit, freeze-dried vegetables, vitamins, minerals, plant and herbal extracts or mixtures thereof. 6. Individualized probiotic composition according to claims 1 to 5, characterized in that the probiotic culture component contains up to 34% by volume of probiotic cultures of the genus Bifidobacterium and 66 to 99% by volume of probiotic cultures of the genus Lactobacillus of the total volume of these cultures in the probiotic culture component. 7. Individualized probiotic composition according to claims 1 to 5, characterized in that the probiotic culture component contains 2 to 49% by volume of probiotic cultures of the genus Bifidobacterium and 51 to 98% by volume of probiotic cultures of the genus Lactobacillus of the total volume of these cultures in the probiotic culture component. 8. Individualized probiotic composition according to claims 1 to 5, characterized in that the probiotic culture component contains 20 to 80% by volume of probiotic cultures of the genus Bifidobacterium and 20 to 80% by volume of probiotic cultures of the genus Lactobacillus of the total volume of these cultures in the probiotic culture component. 9. Individualized probiotic composition according to claims 1 to 5, characterized in that the probiotic culture component contains 20 to 99% by volume of probiotic cultures of the genus Bifidobacterium and 1 to 80% by volume of probiotic cultures of the genus Lactobacillus of the total volume of these cultures in the probiotic culture component. 10. A food, pharmaceutical or veterinary product, characterized in that it contains an individualized probiotic composition according to claims 1 to 9. - 22 CZ 38830 U1 11. A food, pharmaceutical or veterinary product according to claim 10, characterized in that it is in the form of a food supplement, yogurt, kefir, tablet, capsule, powder, gel, suppository, solution, granules or granulate. 12. A cosmetic product, characterized in that it contains an individualized probiotic composition according to claims 1 to 9. 13. A cosmetic product according to claim 12, characterized in that it is in the form of a cream, ointment, body oil, mouthwash, soap or shampoo.