为了得到可抑制口腔致病菌的乳酸菌,从四川大凉山农家泡菜中分离筛选出9株乳酸菌,使用牛津杯法,以变异链球菌、白色念珠菌、具核梭杆菌、牙龈卟啉单胞菌为指示菌,对筛选的菌株进行抑菌试验,随后进行16S rRNA 基因序列鉴定。每隔4 h测定菌株SD26上清液的pH值及对白色念珠菌的抑菌活力,并将不同pH的SD26上清液与白色念珠菌共培养,测定其对白色念珠菌生长过程的影响。将SD26上清液进行酸碱和盐析处理,测定其对白色念珠菌的抑菌活力。最后测定SD26上清液对食源性致病菌的抑菌谱。结果表明,初筛的9株乳酸菌中SD26对白色念珠菌的抑菌活性最高,抑菌圈直径为(26.63±0.30) mm。经鉴定SD26为植物乳杆菌(Lactobacillus plantarum),培养20 h时,上清液pH为3.81,对白色念珠菌的抑菌圈直径最大,且SD26上清液对白色念珠菌抑制作用具有低pH环境的依赖性,盐析中性沉淀复溶液无抑菌活性。SD26上清液对食源性致病菌中单核细胞增生李斯特氏菌 (CMCC54002)的抑菌作用最强,抑菌圈直径为(22.68±0.06) mm。研究表明,SD26上清液中对白色念珠菌起抑菌作用的物质为有机酸,该研究为应用乳酸菌防治口腔致病菌提供了基础。
In order to obtain the lactic acid bacteria that can inhibit oral pathogens, nine strains of lactic acid bacteria were isolated and screened from cottage pickles in Daliangshan, Sichuan. Antibacterial tests of selected isolates were conducted using Oxford cup method, with Streptococcus mutans, Candida albicans, Fusobacterium nucleatum, and Porphyromonas gingivalis as indicator bacteria. Then 16S rRNA gene sequencing was performed to identify the screened lactic acid bacteria. The pH of SD26 supernatant and its antibacterial activity against C. albicans were measured every 4 hours. Further, the SD26 supernatant of different pH was co-cultured with C. albicans to determine its effect on the growth of C. albicans. The SD26 supernatant was treated with acid, base and salt to determine its antibacterial activity against C. albicans. Finally, this study determined the antimicrobial spectrum of SD26 supernatant against foodborne pathogens. The results showed that SD26 had the highest antibacterial activity against C. albicans among the nine isolates, and the diameter of inhibition zone was (26.63±0.30) mm. SD26 was identified as Lactobacillus plantarum. The pH value of the supernatant was 3.81 when cultured for 20 hours, and the diameter of the inhibition zone against C. albicans was the largest. Furthermore, the inhibitory effect of SD26 supernatant on C. albicans depended on low pH environment, and the salting-out neutral precipitation complex solution had no antibacterial activity. SD26 supernatant had the strongest inhibitory effect on Listeria monocytogenes CMCC54002 among foodborne pathogens, with the inhibitory zone diameter of (22.68±0.06) mm. The results indicated that organic acids were the main antibacterial substances against C. albicans in the supernatant of SD26, which lays a foundation for the application of lactic acid bacteria in the prevention and treatment of oral pathogens.
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