Survival of microorganisms from modern probiotics in model conditions of the intestine
Ключові слова:
probiotics, intestine, stability to acids and bile.Анотація
Introduction. The staye of intestinal microflora affects the work of the whole organism. When composition of normal ibtestine microflora changes, its restoration is required. In our days a wide variety of probiotic drugs are available on the market which can be used to solve this problem. Most bacteria having probiotic properties represent the families Lactobacillus and Bifidobacterium, which have poor resistance to acidic content of the stomach and toxic effects of bile salts. Various studies have clearly shown that in a person with normal acidic and bile secretion, the lactobacilli and bifidobacteria are not detected after the passage through the duodenum, i.e., they perish before reaching the small intestines. In this study we compared the survival of different microorganisms which are contained in 9 probiotic drugs in a model of gastric and intestinal environments.Material and methods. In the laboratory of SI: “Mechnikov Institute Microbiology and Immunology, National Ukrainian Academy Medical Sciences" the in vitro experimentshave been evaluated to test the ability of different probiotic bacteria which were contained in 9 probiotic drugs to survive the impact of the model environment of the stomach and duodenum. Bacillus coagulans persistence was evaluated under impact of simulated environment of the stomach and duodenum, it also was assessed by the quantity of CFU by incubation on culture medium. The following were studied: Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus reuteri, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus bulgaricus, Bifidobacterium bifidum, Bifidobacterium longum , Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium animalis subsp. Lactis BB-12, Saccharomyces boulardii, Bacillus coagulans, Bacillus clausii, Enterococcus faecium. Microorganisms were incubated for 3 hours in a model environment of the stomach (pepsin 3 g / l, hydrochloric acid of 160 mmol / l, pH 2.3), later after centrifugation and washing, they were incubated for 3 hours in intestinal model environment (bile salts 3% pancreatin 0.1%, pH 7.0). Inoculation was performed before incubation, after incubation in the gastric medium and after incubation in intestinal medium. We used the medium corresponding to the studied genus of bacteria - MRS-environment for lactobacilli, bifidum for Bifidobacterium, sabouraud medium for the isolation of yeasts and fungi and endo agar for the isolation of Enterobacteriaceae. We assessed the quantity of CFU before and after impact.Results and discussion. After incubation in a simulated gastric environment, bacteria of the type Lactobacillus and Bifidobacterium did not survive and were not defined. Only Bacillus coagulans and Saccharomyces boulardii were resistant. These microorganisms grew after incubation in the same amount as before incubation - 105-6 and 107-8 CFU respectively. Bacillus clausii also survived in these conditions, but to a lesser extent: initially - 107 CFU, after incubation - 105 CFU. After staying in model environment of the duodenum Bacillus coagulans and Saccharomyces boulardii were still fully viable, and the number of germinating Bacillusclausii bacteria decreased by an order - up to 104 CFU.Conclusion. The probiotics containing Bacillus coagulans and Saccharomyces boulardii showed complete resistance to the impact of the model environment of the stomach and duodenum, Bacillus clausii was partially resistant. It leads to conclusion that probiotic drugs containing lactobacilli and bifidobacteria, cannot withstand the aggressive environmental influence of the stomach and duodenum and become inactivated under their influence. Probiotic drugs Enterol containing yeast Saccharomyces boulardii, and Laktovit Forte containing the spore-forming bacterium Bacillus coagulans are completely resistant to the action of the model environment of the stomach and duodenum.
Посилання
Shulpecova YuO. Intestinal bacteria, probiotics and prospects of their use for the treatment of diseases of the gastrointestinal tract. URL: http://medi.ru/doc/1951116.htm
Merritt ME, Donaldson JR. Effect of bile salts on the DNA and membrane integrity of enteric bacteria. // Journal of Medical Microbiology. 2009. № 58. P. 1533-41.
Tennant SM, Hartland EL, Phumoonna T, Lyras D, Rood JI, Robins-Browne RM, Van Driel IR. Influence of gastric acid on susceptibility to infection with ingested bacterial pathogens. // Infect Immun. 2008. № 76. P. 639–45.
Sánchez B, Champomier-Vergès M-C, Collado MC. [et al.] Low-pH Adaptation and the Acid Tolerance Response of Bifidobacterium longum Biotype longum. // Appl Environ Microbiol. 2007. Vol. 73. №. 20. P. 6450–59.
O'May GA, Reynolds N, Smith AR, Kennedy A, Macfarlane GT. Effect of pH and antibiotics on microbial overgrowth in the stomachs and duodena of patients undergoing percutaneous endoscopic gastrostomy feeding. // J. Clin. Microbiol. 2005. № 43. P. 3059-65.
Dixit G, Samarth D, Tale V, Bhadekar R. Comparative studies on potential probiotic characteristics of Lactobacillus acidophilus strains. // Eurasia J Biosci. 2013. № 7. Р. 1-9.
EL-Shafei K, Tawfik NF, Dabiza MA, Sharaf OM, Effat BA. In vitro assessment of gastrointestinal viability of potentially probiotic Lactobacilli // J. Am. Sci. 2010. Vol. 6. №11. Р. 357-367.
Marteau P, Minekus M, Havenaar R, Huis in't Veld JH. Survival of lactic acid bacteria in a dynamic model of the stomach and small intestine: validation and the effects of bile. // J Dairy Sci. 1997. Vol. 80. №6. Р. 1031-37.
Duc LH, Hong HA, Cutting SM. Germination of the spore in the gastrointestinal tract provides a novel route for heterologous antigen presentation. // Vaccine. 2003. № 21. P. 4215–24.
Ripamonti B, Agazzi A, Bald A. [et al.] Administration of Bacillus coagulans in calves: recovery from faecal samples and evaluation of functional aspects of spores. // Vet. Res. Commun. 2009. Vol. 33. № 8. P. 991-1001.
Methodical instructions for the use of unified microbiological (bacteriological) methods of investigation in clinical diagnostic laboratories. Appendix No. 1 to Order No. 535 of the USSR Ministry of Health of 22.04.1985.
##submission.downloads##
Як цитувати
Номер
Розділ
Ліцензія
Авторське право (c) 2019 Annals of Mechnikov's Institute
Ця робота ліцензованаІз Зазначенням Авторства – Некомерційна – Без Похідних 3.0 Міжнародна.