Pulsed-Field Gel Electrophoresis and Lectin Blotting Analyses of Bifidobacterium longum Strains Isolated from Human Feces and Probiotic Products

Juri Hamatsuka, T. Maegawa, Y. Nishitani, R. Osawa
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引用次数: 2

Abstract

Pulsed-field gel electrophoresis and lectin blotting were performed on a total of 41 B. longum strains isolated from 27 human fecal samples and 14 probiotic products in order to characterize their clonality and variation in expression of exopolysacchrides (EPSs; cell-bound polysaccharides). The probiotic isolates formed several distinct clonal clusters, and most of them shared the same lectin blotting profile. Almost all human fecal isolates were distinct from the probiotic isolates not only clonally but also in terms of EPS expression. Most of the fecal isolates within each clonal cluster presented different lectin blotting profiles, suggesting that the structures of EPSs from B. longum were of a labile nature independent of their clonality. Lectin blotting was also performed on a total 38 B. longum strains that had been isolated periodically (at the 1st, 23 rd, and 68th weeks) from fecal samples of 12 human subjects in order to evaluate whether EPS expression was host-specific. B. longum strains that had been isolated from fecal samples of the same individual at different times presented identical or almost identical lectin blotting profiles in 9 out of 12 subjects despite being clonally distant from each other. These findings suggest that variations in EPS expression are related to the hosts but not to the strains' clonality. This in turn suggests that human hosts can discriminate indigenous strains from exogenous ones based on EPS expression profiles.
人类粪便和益生菌制品中长双歧杆菌的脉冲场凝胶电泳和凝集素印迹分析
采用脉冲场凝胶电泳和凝集素印迹技术对从27份人体粪便样本和14种益生菌产品中分离得到的41株长芽孢杆菌进行了克隆性和体外多聚糖(EPSs)表达变化的分析;cell-bound多糖)。分离的益生菌形成了几个不同的克隆簇,其中大多数具有相同的凝集素印迹图谱。几乎所有的人粪便分离株与益生菌分离株不仅在克隆上不同,而且在EPS表达方面也不同。每个克隆簇内的大多数粪便分离株具有不同的凝集素印迹图谱,表明长叶假单胞菌的eps结构具有不稳定的性质,与克隆无关。我们还对从12名受试者的粪便样本中定期(第1周、第23周和第68周)分离的38株长芽孢杆菌进行了凝集素印迹,以评估EPS的表达是否具有宿主特异性。在不同时间从同一个体的粪便样本中分离出的长芽孢杆菌菌株,尽管彼此克隆距离遥远,但在12名受试者中有9名的凝集素印迹图谱相同或几乎相同。这些结果表明,EPS的表达变化与宿主有关,而与菌株的克隆性无关。这反过来表明人类宿主可以根据EPS表达谱区分本地菌株和外源菌株。
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