K Liu1, B-X Hou. 1. Department of Endodontics and Operative Dentistry, School of Stomatology, Capital Medical University, Dongcheng District, Beijing, China. houbenxiang@gmail.com.
Abstract
OBJECTIVE: To evaluate the effect of dltA-deficient mutant on the bacterial growth and biofilm formation by P. micra ATCC 33270. Parvimonas micra contributes to many human polymicrobial infections, and is common in dental plaque biofilms of patients with periodontal and endodontic conditions. Lipoteichoic acid (LTA) performs several functions in gram-positive bacteria, including maintenance of cationic homeostasis and modulation of autolytic activities. The activation of dltA gene expression protects LTA expressing gram-positive bacteria from innate immune anti-microbial defense. MATERIALS AND METHODS: Deficient mutant of the dltA gene was created from P. micra ATCC 33270 by homologous recombination. Colony-forming units (CFUs) and turbidity helped estimate the growth of P. micra. Crystal violet staining, Confocal Scanning Laser Microscopy (CSLM) and Scanning Electron Microscopy (SEM) evaluated biofilm mass and structure. RESULTS: P. micra ATCC 33270 with dltA-deficient mutant was successfully established. CFUs of the wild-type strains were significantly higher than that of the dltA-deficient mutant strains after 24 h, 48 h, 72 h and 7 d culture (all p < 0.05). The growth rate of dltA-deficient mutant strains was significantly lower than their wild-type counterparts. Furthermore, crystal violet staining showed that the dltA mutant formed significantly less biofilm as compared to wild-type strains. The dltA-deficient mutant synthesized a thin and incomplete biofilm after incubation for 48 h. With increasing incubation time, all biofilm units were seen to shrink, and this structure almost disappeared after 7 days of culture as observed by CSLM and SEM. CONCLUSIONS: The dltA gene is associated with bacterial growth and biofilm formation by P. micra ATCC 33270.
OBJECTIVE: To evaluate the effect of dltA-deficient mutant on the bacterial growth and biofilm formation by P. micra ATCC 33270. Parvimonas micra contributes to many humanpolymicrobial infections, and is common in dental plaque biofilms of patients with periodontal and endodontic conditions. Lipoteichoic acid (LTA) performs several functions in gram-positive bacteria, including maintenance of cationic homeostasis and modulation of autolytic activities. The activation of dltA gene expression protects LTA expressing gram-positive bacteria from innate immune anti-microbial defense. MATERIALS AND METHODS: Deficient mutant of the dltA gene was created from P. micra ATCC 33270 by homologous recombination. Colony-forming units (CFUs) and turbidity helped estimate the growth of P. micra. Crystal violet staining, Confocal Scanning Laser Microscopy (CSLM) and Scanning Electron Microscopy (SEM) evaluated biofilm mass and structure. RESULTS:P. micra ATCC 33270 with dltA-deficient mutant was successfully established. CFUs of the wild-type strains were significantly higher than that of the dltA-deficient mutant strains after 24 h, 48 h, 72 h and 7 d culture (all p < 0.05). The growth rate of dltA-deficient mutant strains was significantly lower than their wild-type counterparts. Furthermore, crystal violet staining showed that the dltA mutant formed significantly less biofilm as compared to wild-type strains. The dltA-deficient mutant synthesized a thin and incomplete biofilm after incubation for 48 h. With increasing incubation time, all biofilm units were seen to shrink, and this structure almost disappeared after 7 days of culture as observed by CSLM and SEM. CONCLUSIONS: The dltA gene is associated with bacterial growth and biofilm formation by P. micra ATCC 33270.
Authors: Dustin L Higashi; Sean McGuire; Yasser M Abdelrahman; Zhengzhong Zou; Hua Qin; David Anderson; Elizabeth A Palmer; Jens Kreth; Justin Merritt Journal: Microbiol Spectr Date: 2022-04-13