ИЗУЧЕНИЕ БИОЛОГИЧЕСКИХ СВОЙСТВ АУТОШТАММОВ LACTOBACILLUS SPP. ДЛЯ СОЗДАНИЯ ПРОБИОТИКОВ

Хакимова Л.Р.Ахметова Л.Р.Гимранова И.А.Потапова С.М.

Doi: 10.51620/0869-2084-2023-68-8-480-488 ISSN: 0869-2084 (Print) ISSN: 2412-1320 (Online)

Аннотация Об авторах Список литературы Ключ. слова

Аннотация

Пробиотики представляют собой фармацевтические препараты, БАД или пищевые добавки с живыми бактериями, которые оказывают восстанавливающее влияние на здоровье человека, способствуют активации иммунной системы, улучшают течение аллергических состояний. Часто в качестве пробиотиков, используются бактерии Lactobacillus spp. Правильное употребление пробиотиков может быть эффективным при лечении дисбактериозов, диареи различного генеза, заболеваний полости рта, мочеполовой системы и т. п. Цель исследования: отбор аутоштаммов, которые могут быть использованы в качестве основы при создании пробиотических препаратов. Изучены 182 штамма Lactobacillus spp., из них отобраны девять: 9SM, Dec 1, Ku-f, 12 Pet-f, 14 Ul-d, 15 Sul-c, 10 POD, 7LV, Куз 2, которые идентифицированы до вида и внесены во Всероссийскую коллекцию промышленных микроорганизмов (БРЦ ВКПМ) (Москва). Все отобранные аутоштаммы лактобактерий обладают высоким уровнем перекрёстной адгезии, показали индекс адгезивности микроорганизмов (ИАМ)≥40,0 больше, чем у 50% пациентов. Лучшую адгезию показал аутоштамм 10 POD (ИАМ≥40,0 у 80% исследуемых пациентов). Все исследуемые штаммы Lactobacillus spp. устойчивы к ванкомицину, промежуточную устойчивость к антибиотику норфлоксацину имеют штаммы Ku-f, Куз 2, 15 Sul-c, устойчивыми оказались Dec 1 и 7LV. Исследуемые штаммы активно ингибируют рост E. coli и S. aureus, обладают высокой адгезионной способностью, имеют хорошую кислотообразующую и антагонистическую активности, что указывает на их высокую эффективность в качестве пробиотических бактерий. Показано, что рассматриваемые аутоштаммы Lactobacillus spp. могут быть определены как кандидаты для создания новых эффективных пробиотиков.

Annotation

Probiotics are pharmaceutical preparations, dietary supplements or nutritional supplements with live bacteria that have a restorative effect on human health, contribute to the activation of the immune system, and improve the course of allergic conditions. The bacteria most commonly used as probiotics are Lactobacillus spp. Proper use of probiotics can be effective in the treatment of dysbacteriosis, diarrhea of various origins, diseases of the oral cavity, genitourinary system, etc. The purpose of the study: the selection of autostrains that can be used as a basis for the creation of probiotic preparations. In our study, 182 strains of Lactobacillus spp. were studied, nine of them were selected: 9SM, Dec 1, 12 Pet-f, Ku-f, 14 Ul-d, 15 Sul-c, 10 POD, 7LV, Kuz 2, which were identified to species and included in the All-Russian Collection of Industrial Microorganisms (Moscow). All these autostrains of lactobacilli have a high level of cross-adhesion. They showed an index of microorganism adhesiveness (IAM)≥40.0 greater than in 50% of patients. The 10 POD autostrain showed the best adhesion (IAM≥40.0 in 80% of the studied patients). All studied strains of Lactobacillus spp. resistant to vancomycin, strains Ku-f, Kuz 2, 15 Sul-c have intermediate resistance to the antibiotic norfloxacin, and Dec 1 and 7LV turned out to be resistant. The studied strains actively inhibit the growth of E. coli and S. aureus, have a high adhesive ability, have good acid-forming and antagonistic activities, which indicates their high efficiency as probiotic bacteria. As a result of the research, it was shown that the examined autostrains of Lactobacillus spp. can be identified as candidates for the creation of new effective probiotics.

Об авторах

Список литературы

1. Tochilina A. G., Belova I. V., Solovieva I. V., Novikova N. A., Ivanova T. P. Study of the biological properties of strains of the genus Lactobacillus. Sovremennye problemy nauki i obrazovaniya. 2015; 5:631. (in Russian)
2. Hill C., Guarner F., Reid G., Gibson G. R., Merenstein D. J., Pot B. et al. Expert consensus document. The International scientific association for probiotics and prebiotics cStatement on the scope and appropriate use of the term probiotic. Nat. Rev. Gastroenterol. Hepatol. 2014; 11: 506-14. DOI: 10.1038/nrgastro.2014.66.
3. Chervinets V.M., Mironov A.Yu., Chervinets Yu.V., Bazlov S.N. The state and role of esophagus, stomach, intestinal microbiota in patients with ulcer disease, chronic gastritis, esophagitis. Klinicheskaya Laboratornaya Diagnostika. 2020; 65(1): 42-9. DOI: 10.18821/0869-2084-2020-65-1-42-49. (in Russian)
4. Chervinets Yu.V., Chervinets V.M., Mironov A.Yu. Symbiotic relationship between lactobacilli and microorganisms of the gastrointestinal tract. Tver`: Tsentr Tverskogo gosudarstvennogo meditsivskogo universiteta; 2016. (in Russian)
5. Bonfili L., Cecarini V., Gogoi O., Gong C., Cuccioloni M., Angeletti M., Giacomo Rossi Eleuteri A.M. Microbiota modulation as preventative and therapeutic approach in Alzheimer’s disease. FEBS J. 2021; 288: 2836-55. DOI: 10.1111/febs.15571.
6. Hu J., Wang C., Ye L., Yang W., Huang H., Meng F. et al. Anti-tumour immune effect of oral administration of Lactobacillus plantarum to CT26 tumour-bearing mice. J. Biosci. 2015; 40: 269-79. DOI: 10.1007/s12038-015-9518-4.
7. Kahouli I., Malhotra M., Westfall S., Alaoui-Jamali M. A., Prakash S. Design and validation of an orally administrated active L. fermentum-L. acidophilus probiotic formulation using colorectal cancer Apc Min/+ mouse model. Appl. Microbiol. Biotechnol. 2017; 101: 1999-2019. DOI: 10.1007/s00253-016-7885-x.
8. Waigankar S.S., Patel V. Role of Probiotics in Urogenital Healthcare. J. Midlife Health. 2011; 2: 5-10. DOI: 10.4103/0976-7800.83253.
9. Sunita G., Mallapa R.H., Kumar S.A., Kumar B.V. Probiotics for human health – new innovations and emerging trends. Gut Pathog. 2012; 4: 15. DOI: 10.1186/1757-4749-4-15.
10. Kang J.H., Yun S.I., Park M.H., Park J.H., Jeong S.Y., Park H.O. Anti-obesity effect of Lactobacillus gasseri BNR17 in high-sucrose diet-induced obese mice. PloS One. 2013; 8: e54617. DOI: 10.1371/journal.pone.0054617.
11. Takeda S., Hidaka M., Yoshida H., Takeshita M., Kikuchi Y., Tsend-Ayush C. et al. Antiallergic activity of probiotics from mongolian dairy products on type I allergy in mice and mode of antiallergic action. J. Funct. Foods. 2014; 9: 60-9. DOI: 10.1016/j.jff.2014.04.013.
12. Allaker R.P., Stephen A.S. Use of probiotics and oral health. Curr. Oral. Health Rep. 2017; 4: 309-18. DOI: 10.1007/s40496-017-0159-6.
13. Ince G., Gursoy H., Ipci S.D., Cakar G., Emekli-Alturfan E., Yilmaz S. Clinical and biochemical evaluation of lozenges containing Lactobacillus reuteri as an adjunct to non-surgical periodontal therapy in chronic periodontitis. J. Periodontol. 2015; 86: 746-54. DOI: 10.1902/jop.2015.140612.
14. Chervinets V.M., Chervinets Yu.V., Chichanovskaya L.V., Ganzya D.V., Grigoryants E. O., Belyaev V.S. et al. Spectrum of gas signaling molecules of intestinal lactobacilli in patients with ischemic stroke. Klinicheskaya Laboratornaya Diagnostika. 2022; 67(3): 163-9. DOI: 10.51620/0869-2084-2022-67-3-163-169. (in Russian)
15. Ohshima T., Kojima Y., Seneviratne C.J., Maeda N. Therapeutic application of synbiotics, a fusion of probiotics and prebiotics, and biogenics as a new concept for oral candida infections: a mini review. Front. Microbiol. 2016; 7: 10. DOI: 10.3389/fmicb.2016.00010.
16. Yoo J.I., Shin I.S., Jeon J.G., Yang Y.M., Kim J.G., Lee D.W. The effect of probiotics on halitosis: a systematic review and meta-analysis. Probiotics Antimicrob. Proteins. 2019; 11: 150-7. DOI: 10.1007/s12602-017-9351-1.
17. Sivamaruthi B.S., Kesika P., Chaiyasut C. A review of the role of probiotic supplementation in dental caries. Probiotics Antimicrob. Proteins. 2020; 12: 1300-9. DOI: 10.1007/s12602-020-09652-9.
18. Zhang Y., Ding Y., Guo Q. Probiotic species in the management of periodontal diseases: an overview. Front Cell. Infect. Microbiol. 2022; 12: 806463. DOI: 10.3389/fcimb.2022.806463.
19. Chervinets V.M., Chervinets Yu.V., Leont’eva A.V., Kozlova E.A.,Stulov N.M., Belyaev V.S. et al. Microbiome of the oral cavity in patients with periodontitis, adhesive and biofilm-forming properties. Klinicheskaya Laboratornaya Diagnostika. 2021; 66(1): 45-51. DOI: 10.18821/0869-2084-2021-66-1-45-51. (in Russian)
20. Bafeta A., Koh M., Riveros C., Ravaud P. Harms reporting in randomized controlled trials of interventions aimed at modifying microbiota: a systematic review. Ann. Intern. Med. 2018; 169: 240. DOI: 10.7326/M18-0343.
21. Medical microbiology, virology and immunology: Textbook for students of medical universities. Vorob`yov A.A., Bykov A.S., Boychenko M.N. 3rd ed. Moscow: Meditsinskoe informatsionnoe agentstvo; 2022. ISBN 978-5-9986-0478-2. (in Russian)
22. Il`in V.K., Kiryukhina N.V. Disruption of the colonization resistance syndrome in humans in altered habitats and its prevention. Acta Naturae. 2014; 6(2 (21)): 10-8.
23. Laparra J.M., Sanz Y. Comparison of in vitro models to study bacterial adhesion to the intestinal epithelium. Lett. Appl. Microbiol. 2009; 49: 695-701. DOI: 10.1111/j.1472-765X.2009.02729.x.
24. Collado M.C., Meriluoto J., Salminen S. Role of commercial probiotic strains against human pathogen adhesion to intestinal mucus. Lett. Appl. Microbiol. 2007; 45: 454-60. DOI: 10.1111/j.1472-765X.2007.02212.x.
25. Salminen M.K., Rautelin H. Tynkkynen S. Lactobacillus bacteremia, species identification, and antimicrobial susceptibility of 85 blood isolates. Clin. Infect. Dis. 2006; 42: e35-44. DOI: 10.1086/500214.
26. Fontana L., Bermudez-Brito M., Plaza-Diaz J., Muñoz-Quezada S., Gil A. Sources, isolation, characterisation and evaluation of probiotics. British Journal of Nutrition. 2013; 109(S2): S35-S50. DOI: 10.1017/S0007114512004011.
27. Shenderov B.A. The role of personal functional nutrition in modern anti-aging medicine programs. Vestnik vosstanovitel’noy meditsiny. 2009; 3(31):9-17. (in Russian)
28. Boytsov A.G., Rishchuk S.V., Ilyasov Yu.Yu., Grechaninova T.A. Adhesion of lactobacilli to cells of the vaginal and buccal epithelium. Vestnik Sankt-Peterburgskoy meditsinskoy akademii imeni I.I. Mechnikov. 2004; 4(5):191-3. (in Russian)
29. Sablin O.A., Mikhailov N.V., Yurin M.V., Ilchishina T.A., Kondrashin A.S., Kobiashvili M.G., Zhibrun A.B. Primary resistance of Helicobacter pylori to antibiotics in St. Petersburg. Eksperimental`naya i klinicheskaya gastroenterologiya. 2012; 8:18-23. (in Russian)
30. Jamal M., Ahmad W., Andleeb S., Jalil F., Imran M., Nawaz M.A. et al. Bacterial biofilm and associated infections. J. Chin. Med. Assoc. 2018; 81: 7-11. DOI: 10.1016/j.jcma.2017.07.012.
31. Vasiee A., Falah F., Mortazavi S.A. Evaluation of probiotic potential of autochthonous lactobacilli strains isolated from Zabuli yellow kashk, an Iranian dairy product. J. Appl. Microbiol. 2022; 133: 3201–14. DOI: 10.1111/jam.15772.
32. Barzegari A., Kheyrolahzadeh K., Khatibi S.M.H., Sharifi S., Memar M.Y., Vahed S.Z. The battle of probiotics and their derivatives against biofilms. Infect. Drug. Resistance. 2020; 13: 659-72. DOI: 10.2147/IDR.S232982.
33. Gou H.-Z., Zhang Y.-L., Ren L.-F., Li Z.-J., Zhang L. How do intestinal probiotics restore the intestinal barrier? Front. Microbiol. 2022; 13: 929346. DOI: 10.3389/fmicb.2022.929346.
34. Wang P., Chen S., Liao C., Jia Y., Li J., Shang K. et al. Probiotic properties of chicken-derived highly adherent lactic acid bacteria and inhibition of enteropathogenic bacteria in Caco-2 cells. Microorganisms. 2022; 10(12): 2515. DOI: 10.3390/microorganisms10122515.
35. Mathipa M.G., Thantsha M.S. Probiotic engineering: Towards development of robust probiotic strains with enhanced functional properties and for targeted control of enteric pathogens. Gut. Pathog. 2017; 9: 28. DOI: 10.1186/s13099-017-0178-9.
36. Gueimonde M., Sanchez B.G., de Los Reyes-Gavilán C., Margolles A. Antibiotic resistance in probiotic bacteria. Front Microbiol. 2013; 4:202. DOI: 10.3389/fmicb.2013.00202.
37. Goldstein E.J., Citron D.M., Merriam C.V., Warren Y., Tyrrell K., Fernandez H.T. In vitro activities of dalbavancin and nine comparator agents against anaerobic Gram-positive species and corynebacteria. Antimicrob. Agents Chemother. 2003; 47: 1968-71. DOI: 10.1128/AAC.47.6.1968-1971.2003.
38. Goldstein E.J.C., Tyrrell K.L., Citron D.M. Lactobacillus species: taxonomic complexity and controversial susceptibilities. Clinical Infectious Diseases. 2015; 60(2): 98-S107. DOI: 10.1093/cid/civ072.

Ключевые слова

#Lactobacillus spp.#адгезия #антагонизм #антибиотикочувствительность #аутоштаммы #кислотообразование

Для цитирования: Хакимова Л.Р., Потапова С.М., Ахметова Л.Р., Гимранова И.А. Изучение биологических свойств аутоштаммов Lactobacillus spp. для создания пробиотиков. Клиническая лабораторная диагностика. 2023; 68 (8): 480-488. DOI: https://doi.org/10.51620/0869-2084-2023-68-8-480-488.

For citation: Khakimova L.R., Potapova S.M., Ahmetova L.R., Gimranova I.A. Study of biological properties of Lactobacillus spp. to create probiotics. Klinicheskaya Laboratornaya Diagnostika (Russian Clinical Laboratory Diagnostics). 2023; 68 (8): 480-488 (in Russ.) DOI: https://doi.org/10.51620/0869-2084-2023-68-8-480-488.