Bacteriocin 3xBiotics Presentation

The antibacterial research, from the past 50 years, has focused on meeting the medical needs caused by life-threatening infectious pathogens. The indiscriminate use of synthetic antibiotics has led to the development of many germs that are resistant to them. Also a major problem with the administration of antibiotics in allopathic medicine, either externally or internally, is the creation of dysbiosis which leads to uncontrolled multiplication of pathogens (eg the development of oral/ vaginal candidiasis as a result of antibiotics).

Modification of the intestinal microbiota can lead to many physical and mental illnesses. In addition, synthetic antibiotics have many side effects as well as contraindications.

The advantage of using fermented medicinal plants is that they have a double effect on the pathogen and immunity, increased bioavailability, pre/ pro and postbiotic intake.

That’s why Pro Natura comes to the aid of patients with a new product – Bacteriocin 3xBiotics.

Bacteriocins¹ are small peptides produced by beneficial bacteria (especially LAB), whose property is to destroy other bacteria by acting on them by blocking protein synthesis. Due to their mechanism of action, they represent a potential solution in solving the current crisis (increasing antibiotic resistance²).

Artemisia (Wormwood) is a medicinal plant, widely used in traditional medicine (from vaginal lavage, gastrointestinal disorders, cancers³), which produces secondary metabolites, has direct antimicrobial effects on many resistant pathogenic bacteria, such as: Mycobacterium tuberculosis⁴, Escheria coli⁵, Helicobacter pylory⁶. In particular, one of the bioactive compounds, artemisinin⁷, has effective results against Gram-positive bacteria, and its antibacterial activity is similar to streptomycin (bactericide). Wormwood is also recommended due to its immunomodulatory, antiviral, antifungal, antiparasitic, anti-inflammatory, antioxidant effects⁸.

Turmeric is a panacea, the over 7 thousand studies conducted with this plant, showing countless beneficial effects (Alzheimer’s⁹, pancreatic/ lung/ breast cancer¹⁰, inflammatory diseases – rheumatoid arthritis, osteoarthritis¹¹, skin conditions – Vitiligo¹², Psoriasis¹³). Turmeric is used as a basic ingredient in Asian cuisine, and helps maintain the health of people in areas with poor hygiene.

In Ayurvedic medicine it is used as an antiseptic, antibacterial and anti-inflammatory. Current research suggests positive effects against the genus Salmonella¹⁴, and a strong fungicidal action¹⁵. Curcumin, the main active ingredient in turmeric, can act not only as an antifungal and antibacterial compound, but also as an antiviral compound, inhibiting replication in a wide range of viruses (Zika¹⁶, HSV-1¹⁷, Influenza A¹⁸).

Chemical research has shown the presence of several beneficial compounds, such as flavonoids, ascorbic acid, tocopherols, citric acid, sterols and minerals, in grapefruit seeds. Their efficacy has been reported¹⁹ against over 800 bacterial and viral strains, numerous fungi and a large number of unicellular (Entamoeba histolytica²⁰)/ multicellular (Ascaris lumbricoides²¹) parasites. It has been shown to help inhibit the proliferation of Candida albicans²², a yeast that can affect probiotic bacteria and affect the health of the gastrointestinal tract. Naringenin, a bioactive component of grapefruit seeds, showed gastroprotective activity, especially by inhibiting Helycobacter pylori²³.

Black garlic is obtained from fresh garlic (Allium sativum L.) which has been fermented for a period of time at a controlled high temperature (60–90 ° C) and under controlled high humidity (80–90%). Compared to fresh garlic, black garlic is sweeter, does not release a strong flavor due to its low allicin content, but has increased bioactivity, the phenolic content is 5-8 times higher than that of fresh garlic, so it has an activity much higher antioxidant²⁴. In terms of its medicinal properties, black garlic has been described as antimicrobial, antiseptic, antiviral, antioxidant, immunostimulant, cardioprotective, hypoglycemic and anticancer²⁵. Its properties are related to the bioactive molecules of garlic, such as its organosulfur compounds and non-volatile amino acids (thiosulfinates). These bioactive molecules also include reducing sugars, minerals, saponins and vitamins (A, B and C complex). Black garlic significantly suppresses the growth of the fungus Penicillium citrium²⁶ and also acts on the bacteria Staphylococcus aureus, Escherichia coli, and Bacillus subtilis²⁷.

Cordyceps, considered the mushroom of kings / gods, is the most famous medicinal mushroom being used since antiquity in Tibet. Over the last few years, a series of studies have been carried out to prove its antibacterial and antiviral properties. The results show the existence of the antibacterial polysaccharide extracted from the Cordyceps fungus, which is able to modify the microbial cell permeability²⁸ (Escherichia coli, Staphyloccocus aureus, Bacillus subtilis, Salmonella paratyphi and Pseudomonas aeruginosa²⁹). In addition to the results of antimicrobial activity, cordimin (antifungal peptide) has been isolated, which inhibits the mycelial growth of Bipolaris maydis, Mycosphaerella arachidicola, Rhizoctonia solani and Candida albicans³⁰.

Ganoderma, known as the „mushroom of immortality”, contains over 400³¹ bioactive compounds needed to maintain the body’s health, including triterpenoids, polysaccharides, nucleotides, sterols, fatty acids and proteins / peptides, which have a number of medicinal effects, mainly antimicrobial (Staphylococcus aureus, Proteusbacillus vulgaris, Bacillus subtilis³²), antifungal (Candida albicans³³), antiviral (especially against herpes³⁴ and HIV³⁵), anti-inflammatory, antioxidant, as well as strengthening the immune system³⁶ (activation of macrophages by polysaccharides, induces the release of cytokines that play a role in transmitting information between immune system cells and leukocytes).

Fermentation³⁷ creates the necessary supply of pre / pro / parapro / postbiotics, which ensures both antiviral and antibacterial effects and the normalization of the intestinal microbiota. In traditional fermentation, unlike industrial fermentation, we will find a multitude of bacteriophages with beneficial regulatory effects on the microbiome.

A mixture of fermented plants in SCOBY cultures was also used: Green tea (Camellia sinensis); Neem (Azadirachta Indica); olive leaves (Olea europaea); Bitter cucumber (Momordica Charantia); Mint (Mentha piperita) – cold plant, fights the inflammation that accompanies the infection; Black cumin (Bunium persicum) – strong antioxidant³⁸ and antifungal³⁹ especially inhibitory effect on Fusarium oxysporum; Oregano (Origanum vulgare) – hepatoprotective element⁴⁰, antifungal (Aspergillus niger)⁴¹, antibacterial (Salmonella enteritidis)⁴² and strong antioxidant⁴³; Licorice (Glycyrrhiza glabra).

These are an excellent source of vitamins (A, B1, B6, B9, B12, C, E, K), minerals (calcium, magnesium, potassium, sodium, iron and zinc-antiviral element).

Also, by the presence in the composition of: Propolis which has a strong antiviral effect⁴⁴ (Varicella zoster (VZV), adenovirus type 5 (ADV), HSV1 (Herpex simplex virus)), antimicrobial effect⁴⁵ (Enterococcus faecalis), antifungal⁴⁶ (Cryptococcus neoformans), immunomodulatory, anti-inflammatory effect; Amrita® „food of the gods” – polyfoam pollen fermented in SCOBY consortium, is rich in polyphenols, flavonoids, organic silicon, vitamins, in addition it has a high content of active substances (antioxidants, hydroxy acids, SCFA- butyric acid favors a normal intestinal flora which in turn determines a high immunity); Shilajit & Triphala maintains and promotes a normal intestinal microbiome, Shilajit „mountain blood” the result of fermentation of plant mass at extremely high pressures, over a long period of time is the main source of humic substances (humic acid, fulvic acid)⁴⁷ that play a role in stimulating the system immune (health enhancer)⁴⁸, strong antioxidant⁴⁹, antiviral (decreases HIV infection and replication, inhibits viral fusion between T44 cells)⁵⁰ and improves cognitive processes⁵¹; The acetic acid resulting from the fermentations ensures the inhibition of the formation of the biofilm necessary for the development of pathogens (Streptococcus pyogenes, Pseudomonas, etc.).

In the composition of Bacteriocin 3xBiotics we find many active principles that have as main action the inhibition of bacterial activity, viral (recent studies at Rockeffeller University have shown that there are 3 bacteria in the intestinal microbiome whose metabolites, isopentenyl adenosine, pyrazine, 5-hydroxytryptamine, have the effect of blocking more than 90% of SARS-CoV-2⁵²) and fungal infection.

Bacteriocin 3xBiotics becomes an ideal therapeutic option for patients who have intolerances, allergies to the administration of classic antibiotics or who have developed resistance to them.

Bibliographical references

1. Yang, S. C., Lin, C. H., Sung, C. T., & Fang, J. Y. (2014). Antibacterial activities of bacteriocins: application in foods and pharmaceuticals. Frontiers in microbiology, 5, 241. https://doi.org/10.3389/fmicb.2014.00241
2. Ventola C. L. (2015). The antibiotic resistance crisis: part 1: causes and threats. P & T : a peer-reviewed journal for formulary management, 40(4), 277–283. The Antibiotic Resistance Crisis (nih.gov)
3. Willcox, M. (2009). Artemisia species: from traditional medicines to modern antimalarials—and back again. The Journal of Alternative and Complementary Medicine, 15(2), 101-109. https://doi.org/10.1089/acm.2008.0327
4. Martini, M. C., Zhang, T., Williams, J. T., Abramovitch, R. B., Weathers, P. J., & Shell, S. S. (2020). Artemisia annua and Artemisia afra extracts exhibit strong bactericidal activity against Mycobacterium tuberculosis. Journal of Ethnopharmacology, 262, 113191. https://doi.org/10.1016/j.jep.2020.113191
5. Mathlouthi, A., Saadaoui, N., & Ben-Attia, M. (2021). Essential oils from Artemisia species inhibit biofilm formation and the virulence of Escherichia coli EPEC 2348/69. Biofouling, 37(2), 174-183. DOI:1080/08927014.2021.1886278
6. El-Sayed, M. A., BaAbbad, R., Balash, A., Al-Hemdan, N. A., & Softah, A. (2013). The potential anti Helicobacter pylori and antioxidant effects of Artemisia judaica. Functional Foods in Health and Disease, 3(9), 332-340. DOI: 31989/ffhd.v3i9.42
7. Suganthi Appalasamy, Kiah Yann Lo, Song Jin Ch’ng, Ku Nornadia, Ahmad Sofiman Othman, Lai-Keng Chan, „Antimicrobial Activity of Artemisinin and Precursor Derived from In Vitro Plantlets of Artemisia annua„, BioMed Research International, vol. 2014, Article ID 215872, 6 pages, 2014. https://doi.org/10.1155/2014/215872
8. Shakya, A. K. (2016). Medicinal plants: Future source of new drugs. International Journal of Herbal Medicine, 4(4), 59-64. DOI:13140/RG.2.1.1395.6085
9. Hamaguchi, T., Ono, K., & Yamada, M. (2010). Curcumin and Alzheimer’s disease. CNS neuroscience & therapeutics, 16(5), 285-297. https://doi.org/10.1111/j.1755-5949.2010.00147.x
10. Gupta, S. C., Patchva, S., & Aggarwal, B. B. (2013). Therapeutic roles of curcumin: lessons learned from clinical trials. The AAPS journal, 15(1), 195–218. https://doi.org/10.1208/s12248-012-9432-8
11. Henrotin, Y., Priem, F. & Mobasheri, A. Curcumin: a new paradigm and therapeutic opportunity for the treatment of osteoarthritis: curcumin for osteoarthritis management. SpringerPlus 2, 56 (2013). https://doi.org/10.1186/2193-1801-2-56
12. Gianfaldoni, S., Wollina, U., Tirant, M., Tchernev, G., Lotti, J., Satolli, F., Rovesti, M., França, K., & Lotti, T. (2018). Herbal Compounds for the Treatment of Vitiligo: A Review. Open access Macedonian journal of medical sciences, 6(1), 203–207. https://doi.org/10.3889/oamjms.2018.048
13. Nardo, V. D., Gianfaldoni, S., Tchernev, G., Wollina, U., Barygina, V., Lotti, J., Daaboul, F., & Lotti, T. (2018). Use of Curcumin in Psoriasis. Open access Macedonian journal of medical sciences, 6(1), 218–220. https://doi.org/10.3889/oamjms.2018.055
14. Sandhya A. Marathe, Rupesh Kumar, Parthasarathi Ajitkumar, Valakunja Nagaraja, Dipshikha Chakravortty, Curcumin reduces the antimicrobial activity of ciprofloxacin against Salmonella Typhimurium and Salmonella Typhi, Journal of Antimicrobial Chemotherapy, Volume 68, Issue 1, January 2013, Pages 139–152, https://doi.org/10.1093/jac/dks375
15. V. B. Martins, D. L. da Silva, A. T. M. Neres, T. F. F. Magalhães, G. A. Watanabe, L. V. Modolo, A. A. Sabino, Â. de Fátima, M. A. de Resende, Curcumin as a promising antifungal of clinical interest, Journal of Antimicrobial Chemotherapy, Volume 63, Issue 2, February 2009, Pages 337–339, https://doi.org/10.1093/jac/dkn488
16. Mounce, B. C., Cesaro, T., Carrau, L., Vallet, T., & Vignuzzi, M. (2017). Curcumin inhibits Zika and chikungunya virus infection by inhibiting cell binding. Antiviral research, 142, 148-157. https://doi.org/10.1016/j.antiviral.2017.03.014
17. Kutluay, S. B., Doroghazi, J., Roemer, M. E., & Triezenberg, S. J. (2008). Curcumin inhibits herpes simplex virus immediate-early gene expression by a mechanism independent of p300/CBP histone acetyltransferase activity. Virology, 373(2), 239-247. https://doi.org/10.1016/j.virol.2007.11.028
18. Chen, T. Y., Chen, D. Y., Wen, H. W., Ou, J. L., Chiou, S. S., Chen, J. M., … & Hsu, W. L. (2013). Inhibition of enveloped viruses infectivity by curcumin. PloS one, 8(5), e62482. https://doi.org/10.1371/journal.pone.0062482
19. English, J. Candida Yeast Protection Program. https://nutritionreview.org/2013/04/candida-yeast-protection-program-part2/
20. Swicegood, C. Mother Nature’s Gift. AFA Watchbird, 28(2), 27-28. https://journals.tdl.org/watchbird/index.php/watchbird/article/view/1635
21. Kothari, V. (2013). Bioactive compounds from plant seeds/seed extracts. Introduction to functional food science, 1st edn. Food Science Publisher, Texas, 86-93. (PDF) Bioactive compounds from plant seeds / seed extracts (researchgate.net)
22. Tsutsumi-Arai, C., Takakusaki, K., Arai, Y., Terada-Ito, C., Takebe, Y., Imamura, T., … & Satomura, K. (2019). Grapefruit seed extract effectively inhibits the Candida albicans biofilms development on polymethyl methacrylate denture-base resin. PloS one, 14(5), e0217496. https://doi.org/10.1371/journal.pone.0217496
23. Zayachkivska, O. S., Konturek, S. J., Drozdowicz, D., Konturek, P. C., Brzozowski, T., & Ghegotsky, M. R. (2005). Gastroprotective effects of flavonoids in plant extracts. Journal of Physiology and Pharmacology. Supplement, 56(1), 219-231. https://agro.icm.edu.pl/agro/element/bwmeta1.element.agro-article-25fb8a4c-a050-41eb-b0bd-9ca34f9572e0
24. Wang, W., & Sun, Y. (2017). In vitro and in vivo antioxidant activities of polyphenol extracted from black garlic. Food Science and Technology, 37, 681-685. https://doi.org/10.1590/1678-457X.30816
25. Tran, G. B., Pham, T. V., & Trinh, N. N. (2019). Black garlic and its therapeutic benefits. Medicinal Plants-Use in Prevention and Treatment of Diseases. Black Garlic and Its Therapeutic Benefits | IntechOpen
26. Fratianni F, Riccardi R, Spigno P, Ombra MN, Cozzolino A, Tremonte P, Coppola R, Nazzaro F. Biochemical Characterization and Antimicrobial and Antifungal Activity of Two Endemic Varieties of Garlic (Allium sativum L.) of the Campania Region, Southern Italy. J Med Food. 2016 Jul;19(7):686-91. DOI: 1089/jmf.2016.0027
27. Guo, Y. (2014). Experimental Study On The Optimization Of Extraction Process Of Garlic Oiland Its Antibacterial Effects. African Journal of Traditional, Complementary and Alternative Medicines, 11(2), 411-414. DOI: 4314/ajtcam.v11i2.27
28. Zhang, Y., Wu, Y. T., Zheng, W., Han, X. X., Jiang, Y. H., Hu, P. L., … & Shi, L. E. (2017). The antibacterial activity and antibacterial mechanism of a polysaccharide from Cordyceps cicadae. Journal of functional foods, 38, 273-279. https://doi.org/10.1016/j.jff.2017.09.047
29. Duarte, N., Ferreira, M. J. U., Martins, M., Viveiros, M., & Amaral, L. (2007). Antibacterial activity of ergosterol peroxide against Mycobacterium tuberculosis: dependence upon system and medium employed. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives, 21(7), 601-604. https://doi.org/10.1002/ptr.2119
30. Wong, J. H., Ng, T. B., Wang, H., Sze, S. C. W., Zhang, K. Y., Li, Q., & Lu, X. (2011). Cordymin, an antifungal peptide from the medicinal fungus Cordyceps militaris. Phytomedicine, 18(5), 387-392. https://doi.org/10.1016/j.phymed.2010.07.010
31. Li, Z., Shi, Y., Zhang, X., Xu, J., Wang, H., Zhao, L., & Wang, Y. (2020). Screening immunoactive compounds of Ganoderma lucidum spores by mass spectrometry molecular networking combined with in vivo zebrafish assays. Frontiers in pharmacology, 11, 287. https://doi.org/10.3389/fphar.2020.00287
32. Li, W. J., Nie, S. P., Liu, X. Z., Zhang, H., Yang, Y., Yu, Q., & Xie, M. Y. (2012). Antimicrobial properties, antioxidant activity and cytotoxicity of ethanol-soluble acidic components from Ganoderma atrum. Food and Chemical Toxicology, 50(3-4), 689-694. https://doi.org/10.1016/j.fct.2011.12.011
33. Nayak, A., Nayak, R. N., & Bhat, K. (2010). Antifungal activity of a toothpaste containing Ganoderma lucidum against Candida albicans-an in vitro study. J Int Oral Health, 2(2), 51-57. http://listamester.hu/store/5207/ganozhi_toothpaste.pdfangol.pdf
34. Eo, S. K., Kim, Y. S., Lee, C. K., & Han, S. S. (1999). Antiherpetic activities of various protein bound polysaccharides isolated from Ganoderma lucidum. Journal of ethnopharmacology, 68(1-3), 175-181. https://doi.org/10.1016/S0378-8741(99)00086-0
35. El-Mekkawy, S., Meselhy, M. R., Nakamura, N., Tezuka, Y., Hattori, M., Kakiuchi, N., … & Otake, T. (1998). Anti-HIV-1 and anti-HIV-1-protease substances from Ganoderma lucidum. Phytochemistry, 49(6), 1651-1657. https://doi.org/10.1016/S0031-9422(98)00254-4
36. Ren, L., Zhang, J., & Zhang, T. (2021). Immunomodulatory activities of polysaccharides from Ganoderma on immune effector cells. Food Chemistry, 340, 127933. https://doi.org/10.1016/j.foodchem.2020.127933
37. Rathee, S., Rathee, D., Rathee, D., Kumar, V., & Rathee, P. (2012). Mushrooms as therapeutic agents. Revista Brasileira de Farmacognosia, 22, 459-474. https://doi.org/10.1590/S0102-695X2011005000195
38. Shahsavari, N., Barzegar, M., Sahari, M.A. et al.Antioxidant Activity and Chemical Characterization of Essential Oil of Bunium persicum . Plant Foods Hum Nutr 63, 183–188 (2008). https://doi.org/10.1007/s11130-008-0091-y
39. Sekine, T., Sugano, M., Majid, A., & Fujii, Y. (2007). Antifungal effects of volatile compounds from black zira (Bunium persicum) and other spices and herbs. Journal of chemical ecology, 33(11), 2123-2132. DOI:1007/s10886-007-9374-2
40. Oniga, I., Pușcaș, C., Silaghi-Dumitrescu, R., Olah, N. K., Sevastre, B., Marica, R., … & Hanganu, D. (2018). Origanum vulgare ssp. vulgare: Chemical composition and biological studies. Molecules, 23(8), 2077. https://doi.org/10.3390/molecules23082077
41. Bedoya-Serna, C. M., Dacanal, G. C., Fernandes, A. M., & Pinho, S. C. (2018). Antifungal activity of nanoemulsions encapsulating oregano (Origanum vulgare) essential oil: in vitro study and application in Minas Padrão cheese. Brazilian journal of microbiology, 49, 929-935. https://doi.org/10.1016/j.bjm.2018.05.004
42. Benedec D, Vlase L, Oniga I, Mot AC, Damian G, Hanganu D, Duma M, Silaghi-Dumitrescu R. Polyphenolic composition, antioxidant and antibacterial activities for two Romanian subspecies of Achillea distans Waldst. et Kit. ex Willd. Molecules. 2013 Jul 24;18(8):8725-39. DOI: 3390/molecules18088725
43. Tusevski, Oliver, Kostovska, Aneta, Iloska, Ana, Trajkovska, Ljubica and Simic, Sonja. „Phenolic production and antioxidant properties of some Macedonian medicinal plants” Open Life Sciences, vol. 9, no. 9, 2014, pp. 888-900. https://doi.org/10.2478/s11535-014-0322-1
44. Hazem, A., Pitică-Aldea, I. M., Popescu, C., Matei, L., Dragu, D., Economescu, M., … & Lupuliasa, D. (2017). The antiviral/virucidal effects of alcoholic and aqueous extracts with propolis. Farmacia, 65(6), 868-76. https://farmaciajournal.com/wp-content/uploads/2017-06-art-08-Hazem_Popescu_Lupuliasa_868-876.pdf
45. Fernandes, F. H., Guterres, Z. D. R., Violante, I. M., Lopes, T. F., Garcez, W. S., & Garcez, F. R. (2015). Evaluation of mutagenic and antimicrobial properties of brown propolis essential oil from the Brazilian Cerrado biome. Toxicology reports, 2, 1482-1488. https://doi.org/10.1016/j.toxrep.2015.11.007
46. Peter, C. M., Waller, S. B., Picoli, T., Osório, L. G., Zani, J. L., Meireles, M. C. A., … & Fischer, G. (2019). Chemical and cytotoxic analyses of three varieties of Brazilian propolis (green propolis, jataí propolis and brown propolis) and its anti-Sporothrix brasiliensis in vitro activity. Arquivo Brasileiro de Medicina Veterinária e Zootecnia, 71, 819-827. https://doi.org/10.1590/1678-4162-9918
47. Carrasco-Gallardo, C., Guzmán, L., & Maccioni, R. B. (2012). Shilajit: a natural phytocomplex with potential procognitive activity. International Journal of Alzheimer’s disease, 2012. https://doi.org/10.1155/2012/674142
48. Meena, H., Pandey, H. K., Arya, M. C., & Ahmed, Z. (2010). Shilajit: A panacea for high-altitude problems. International journal of Ayurveda research, 1(1), 37–40. https://doi.org/10.4103/0974-7788.59942
49. Stohs, S. J., Singh, K., Das, A., Roy, S., & Sen, C. K. (2017). Energy and Health Benefits of Shilajit. In Sustained Energy for Enhanced Human Functions and Activity(pp. 187-204). Academic Press. https://doi.org/10.1016/B978-0-12-805413-0.00012-0
50. Pant, K., Singh, B., & Thakur, N. (2012). Shilajit: a humic matter panacea for cancer. http://14.139.206.50:8080/jspui/bitstream/1/4138/1/shilajit-a-panacea-for-cancer.pdf
51. Carrasco-Gallardo, C., Farías, G. A., Fuentes, P., Crespo, F., & Maccioni, R. B. (2012). Can nutraceuticals prevent Alzheimer’s disease? Potential therapeutic role of a formulation containing shilajit and complex B vitamins. Archives of medical research, 43(8), 699-704. https://doi.org/10.1016/j.arcmed.2012.10.010
52. Piscotta, F. J., Hoffmann, H. H., Choi, Y. J., Small, G. I., Ashbrook, A. W., Koirala, B., Campbell, E. A., Darst, S. A., Rice, C. M., & Brady, S. F. (2021). Metabolites with SARS-CoV-2 Inhibitory Activity Identified from Human Microbiome Commensals. mSphere, 6(6), e0071121. https://doi.org/10.1128/mSphere.00711-21