Gut Microbiology & Immunology Group, Chemistry of Natural and Microbial Products Dept., Pharmaceutical Industries Div., National Research Centre, Cairo, Egypt.
Microbiology and Immunology Department, Faculty of Pharmacy, Badr University in Cairo, Cairo, Egypt.
PLoS One. 2021 Jul 26;16(7):e0255092. doi: 10.1371/journal.pone.0255092. eCollection 2021.
Our aim was to isolate, identify and characterize probiotic bacteria as vitamin producers in particular B2 and B9. 150 human fecal samples were collected and used for isolation of vitamin producers-probiotics. 49 isolates were chosen for screening their genome by PCR for the presence of riboflavin and folic acid genes. As a result, three isolates were selected and their production of the B2 and B9 were confirmed by HPLC. The three isolates were identified on species level by sequencing their 16S rRNA gene which showed 100% identical to strains of Pediococcus acidilactici. Thus, they were named as P. acidilactici WNYM01, P. acidilactici WNYM02, P. acidilactici WNYM03 and submitted to the Genbank database with accession numbers. They met the probiotic criteria by expressing 90-95% survival rate at pH (2.0-9.0) and bile salt up to 2% for 3 h in addition to their antimicrobial activity against gram positive and negative microorganisms. They also showed no hemolytic activity and common pattern for antibiotic susceptibility. Our three strains were tested individually or in mixture in vivo on rat colitis model compared to ulcerative group. The strains were administrated orally to rats in daily dose containing CFU 109 for 14 days then followed by induction of colitis using acetic acid then the oral administration was continued for more four days. The histology results, the anti-inflammatory and anti-oxidative stress biomarkers showed the protective role of the strains compared to the ulcerative group. As a conclusion, we introduce novel three probiotic candidates for pharmaceutical preparations and health applications.
我们的目的是分离、鉴定和表征益生菌作为维生素特别是 B2 和 B9 的生产者。收集了 150 个人类粪便样本,用于分离维生素生产者-益生菌。选择了 49 个分离株,通过 PCR 筛选其基因组中是否存在核黄素和叶酸基因。结果,选择了三个分离株,并通过 HPLC 确认了它们生产 B2 和 B9 的能力。通过测序 16S rRNA 基因鉴定这三个分离株的种水平,其结果显示与乳球菌属(Pediococcus)的菌株完全一致。因此,它们被命名为嗜酸乳球菌(Pediococcus acidilactici)WNYM01、WNYM02 和 WNYM03,并提交给 Genbank 数据库,登录号为。它们通过在 pH(2.0-9.0)和胆汁盐高达 2%的条件下 3 小时内表达 90-95%的存活率,以及对革兰氏阳性和阴性微生物的抗菌活性,符合益生菌标准。它们还没有溶血活性和抗生素敏感性的常见模式。我们的三个菌株在体内单独或混合在大鼠结肠炎模型中进行了测试,与溃疡性结肠炎组进行了比较。这些菌株以每天包含 CFU 109 的剂量口服给予大鼠,持续 14 天,然后用乙酸诱导结肠炎,然后继续口服 4 天。组织学结果、抗炎和抗氧化应激生物标志物显示了这些菌株与溃疡性结肠炎组相比的保护作用。总之,我们介绍了三种新型益生菌候选药物,可用于药物制剂和健康应用。