Bubnov Rostyslav V, Babenko Lidiia P, Lazarenko Liudmyla M, Mokrozub Victoria V, Spivak Mykola Ya
1Zabolotny Institute of Microbiology and Virology, National Academy of Sciences of Ukraine, Zabolotny str., 154, Kyiv, 03143 Ukraine.
Clinical Hospital 'Pheophania' of State Affairs Department, Zabolotny str., 21, Kyiv, 03143 Ukraine.
EPMA J. 2018 Apr 13;9(2):205-223. doi: 10.1007/s13167-018-0132-z. eCollection 2018 Jun.
Probiotics have tremendous potential to develop healthy diets, treatment, and prevention. Investigation of in vitro cultural properties of health-promoting microorganisms like lactic acid bacteria (LAB) and bifidobacteria is crucial to select probiotic strains for treatments based on gut microbiota modulation to justify individualized and personalized approach for nutrition and prevention of variety of diseases. The was to study the biological properties of LAB and bifidobacteria probiotic strains, namely adhesive properties; resistance to antibiotics; and biological fluids (gastric juice, bile, pancreatic enzymes), and to overview the literature in the field.
We studied six LAB strains ( ІМV В-7279, ІМV В-7280, subsp. ІМV В-7281, LB-3 VK6, LE VK8, LM VK7), and two bifidobacteria strains ( VKL, VKB). We characterized tinctorial, culturally morphological, physiological, and biochemical properties of probiotic strains of LAB and bifidobacteria by commonly used research methods. Determination of the resistance to antibiotics was carried out using disc-diffusion method. The effects of gastric juice, bile, and pancreatin on the viability of LAB and bifidobacteria were evaluated. Adhesive properties of LAB and bifidobacteria to epithelial cells were assessed calculating three indicators: average adhesion rate (AAR), participation rate of epithelial cells (PRE), and adhesiveness index of microorganisms (AIM). Electron microscopy of LAB and bifidobacteria cells was conducted.
The studied strains of LAB and bifidobacteria did not form spores, were positively stained by Gram, grow on medium in a wide range of pH (1.0-9.0, optimum pH 5.5-6.5), were sensitive to a wide range of antibiotics; and showed different resistance to gastric juice, bile, and pancreatic enzymes. The most resistant to antibiotics were LB-3 VK6 and LE VK8 strains. The most susceptible to gastric juice was LM VK7, which stopped its growth at 8% of gastric juice; IMV B-7279, VKL, and VKB strains were resistant even in the 100% concentration. Strains IMV В-7279, IMV В-7280, VKL, VKB, LB-3 VK6, LE VK8, and subsp. IMV В-7281 were resistant to pancreatic enzymes. Adhesive properties of the strains according to AIM index were high in IMV В-7280, VKL, and VKB; were moderate in subsp. IMV В-7281; and were low in IMV В-7279, LB-3 VK6, LE VK8, and LM VK7.
We recognized strain-dependent properties of studied LAB and bifidobacteria probiotic strains (adhesive ability, resistance to antibiotics, and gut biological fluids) and discussed potential for most effective individualized treatment for gut and distant sites microbiome modulation.
益生菌在开发健康饮食、治疗和预防方面具有巨大潜力。研究乳酸菌（LAB）和双歧杆菌等促进健康的微生物的体外培养特性，对于选择基于肠道微生物群调节的益生菌菌株进行治疗至关重要，从而为营养和多种疾病的预防提供个性化方法。本研究旨在研究LAB和双歧杆菌益生菌菌株的生物学特性，即黏附特性、对抗生素的抗性以及对生物流体（胃液、胆汁、胰酶）的抗性，并综述该领域的文献。
我们研究了6株LAB菌株（IMV B-7279、IMV B-7280、嗜酸乳杆菌亚种IMV B-7281、植物乳杆菌LB-3 VK6、干酪乳杆菌LE VK8、瑞士乳杆菌LM VK7）和2株双歧杆菌菌株（青春双歧杆菌VKL、两歧双歧杆菌VKB）。我们通过常用的研究方法对LAB和双歧杆菌益生菌菌株的染色、培养形态、生理和生化特性进行了表征。使用纸片扩散法测定对抗生素的抗性。评估了胃液、胆汁和胰蛋白酶对LAB和双歧杆菌活力的影响。通过计算三个指标评估LAB和双歧杆菌对上皮细胞的黏附特性：平均黏附率（AAR）、上皮细胞参与率（PRE）和微生物黏附指数（AIM）。对LAB和双歧杆菌细胞进行了电子显微镜检查。
所研究的LAB和双歧杆菌菌株不形成孢子，革兰氏染色呈阳性，在广泛的pH范围（1.0 - 9.0，最佳pH 5.5 - 6.5）的培养基上生长，对多种抗生素敏感；并且对胃液、胆汁和胰酶表现出不同的抗性。对抗生素最具抗性的是植物乳杆菌LB-3 VK6和干酪乳杆菌LE VK8菌株。对胃液最敏感的是瑞士乳杆菌LM VK7，在8%的胃液中停止生长；IMV B-7279、青春双歧杆菌VKL和两歧双歧杆菌VKB菌株即使在100%浓度下也具有抗性。IMV B-7279、IMV B-7280、青春双歧杆菌VKL、两歧双歧杆菌VKB、植物乳杆菌LB-3 VK6、干酪乳杆菌LE VK8和嗜酸乳杆菌亚种IMV B-7281菌株对胰酶具有抗性。根据AIM指数评估这些菌株的黏附特性，IMV B-7280、青春双歧杆菌VKL和两歧双歧杆菌VKB菌株较高；嗜酸乳杆菌亚种IMV B-7281菌株中等；IMV B-7279、植物乳杆菌LB-3 VK6、干酪乳杆菌LE VK8和瑞士乳杆菌LM VK7菌株较低。
我们认识到所研究的LAB和双歧杆菌益生菌菌株具有菌株依赖性特性（黏附能力、对抗生素的抗性以及对肠道生物流体的抗性），并讨论了对肠道和远处部位微生物群进行最有效个性化治疗的潜力。
