Laboratory of Research in Biofilms Rosário Oliveira (LIBRO), Centre of Biological Engineering (CEB), University of Minho, Braga, Portugal.
Front Cell Infect Microbiol. 2021 Nov 15;11:771666. doi: 10.3389/fcimb.2021.771666. eCollection 2021.
biofilm cells can enter a physiological state known as viable but non-culturable (VBNC), where, despite being alive, they do not grow in conventional laboratory media. As such, the presence of VBNC cells impacts the diagnosis of biofilm-associated infections. Previous transcriptomics analysis of strain 9142 biofilms with higher proportions of VBNC cells suggested that the genes and could be involved in the induction of the VBNC state. However, it was previously demonstrated that VBNC induction is strain-dependent. To properly assess the role of these genes in VBNC induction, the construction of mutant strains is necessary. Thus, herein, we assessed if VBNC cells could be induced in strain 1457, a strain amenable to genetic manipulation, and if the previously identified genes were involved in the modulation of the VBNC state in this strain. Furthermore, we evaluated the formation of VBNC cells on planktonic cultures. Our results showed that despite being commonly associated with biofilms, the proportion of VBNC cells can be modulated in both biofilm and planktonic cultures and that the expression of and was upregulated under VBNC inducing conditions in both phenotypes. Overall, our study revealed that the formation of VBNC cells in is independent of the mode of growth and that the genes and seem to be relevant for the regulation of this physiological condition.
生物膜细胞可以进入一种被称为“活的但非可培养的”(VBNC)的生理状态,在这种状态下,尽管它们还活着,但在常规实验室培养基中却无法生长。因此,VBNC 细胞的存在会影响生物膜相关感染的诊断。先前对含有较高比例 VBNC 细胞的 9142 生物膜进行的转录组学分析表明,基因 和 可能参与诱导 VBNC 状态。然而,先前的研究表明,VBNC 的诱导是菌株依赖性的。为了正确评估这些基因在 VBNC 诱导中的作用,需要构建突变菌株。因此,在此,我们评估了 1457 菌株(一种可进行遗传操作的菌株)是否可以诱导 VBNC 细胞,以及先前鉴定的基因是否参与了该菌株 VBNC 状态的调节。此外,我们还评估了浮游培养物中 VBNC 细胞的形成。我们的结果表明,尽管与生物膜通常相关,但 VBNC 细胞的比例可以在生物膜和浮游培养物中调节,并且在两种表型下,VBNC 诱导条件下 和 的表达均上调。总体而言,我们的研究表明, 形成 VBNC 细胞的方式独立于生长方式,并且基因 和 似乎与这种生理状态的调节有关。