Contribution of Drugs Interfering with Protein and Cell Wall Synthesis to the Persistence of Pseudomonas aeruginosa Biofilms: An In Vitro Model

The occurrence of Pseudomonas aeruginosa (PA) persisters, including viable but non-culturable (VBNC) forms, subpopulations of tolerant cells that can survive high antibiotic doses, is the main reason for PA lung infections failed eradication and recurrence in Cystic Fibrosis (CF) patients, subjected to life-long, cyclic antibiotic treatments. In this paper, we investigated the role of subinhibitory concentrations of different anti-pseudomonas antibiotics in the maintenance of persistent (including VBNC) PA cells in in vitro biofilms. Persisters were firstly selected by exposure to high doses of antibiotics and their abundance over time evaluated, using a combination of cultural, qPCR and flow cytometry assays. Two engineered GFP-producing PA strains were used. The obtained results demonstrated a major involvement of tobramycin and bacterial cell wall-targeting antibiotics in the resilience to starvation of VBNC forms, while the presence of ciprofloxacin and ceftazidime/avibactam lead to their complete loss. Moreover, a positive correlation between tobramycin exposure, biofilm production and c-di-GMP levels was observed. The presented data could allow a deeper understanding of bacterial population dynamics during the treatment of recurrent PA infections and provide a reliable evaluation of the real efficacy of the antibiotic treatments against the bacterial population within the CF lung.     

Molecular dynamics simulations of the interaction between OH radicals in plasma with poly-β-1–6-N-acetylglucosamine

Cold atmospheric plasma shows a satisfactory ability to inactivate bacterial biofilms that are difficult to remove using conventional methods in some cases. However, the researches on the inactivation mechanism are not quite sufficient. Poly-β-1–6-N-acetylglucosamine (PNAG), which is one of the important components in some biofilms, was used as the research subject, and the related mechanism of action triggered by different concentrations of the OH in plasma was studied using reactive molecular dynamics simulations. The results showed that OH radicals could not only trigger the hydrogen abstraction reaction leading to cleavage of the PNAG molecular structure, but undergo an OH addition reaction with PNAG molecules. New reaction pathways appeared in the simulations as the OH concentration increased, but the reaction efficiency first increased and then decreased. The simulation study in this paper could, to some extent, help elucidate the microscopic mechanism of the interaction between OH radicals in plasma and bacterial biofilms at the atomic level.     

次氯酸钠对德尔卑沙门氏菌生物被膜的抑制作用及机制

目的：为控制德尔卑沙门氏菌（Salmonella Derby）在食品加工过程中的污染，研究不同体积分数次氯酸钠对S. Derby生物被膜的抑制作用及其机制。方法：采用结晶紫染色法和平板菌落计数法观察次氯酸钠对生物被膜的抑制和清除效果，进一步观察不同体积分数次氯酸钠作用下沙门氏菌泳动能力和生物被膜内细胞的代谢活性、胞外聚合物含量和微观形态的变化规律。结果：次氯酸钠的最小抑菌浓度（minimum inhibitory concentration，MIC）为0.08%（体积分数，下同），亚MIC（0.04%、0.02%、0.01%）的次氯酸钠可以抑制S. Derby生物被膜的形成，对成熟生物被膜也有一定的清除效果，且不同亚MIC次氯酸钠的作用效果差异显著（P＜0.05）；不同体积分数次氯酸钠处理能降低生物被膜内细胞代谢活性，抑制胞外多糖的合成，且不同体积分数次氯酸钠的作用效果差异显著（P＜0.05）。激光共聚焦扫描显微镜结果显示，与对照组相比，次氯酸钠处理使得生物被膜厚度减小、活菌数减少、死菌数增多、细菌由聚集状态变为分散状态。结论：不同体积分数次氯酸钠对S. Derby生物被膜的形成有明显的抑制作用，这一作用与干扰细菌生长、生物被膜内细胞代谢活性和胞外多糖的合成有关。     

The Antimicrobial Peptide Temporin G: Anti-Biofilm,Anti-Persister Activities,and Potentiator Effect of Tobramycin Efficacy Against Staphylococcus aureus

Bacterial biofilms are a serious threat for human health, and the Gram-positive bacterium Staphylococcus aureus is one of the microorganisms that can easily switch from a planktonic to a sessile lifestyle, providing protection from a large variety of adverse environmental conditions. Dormant non-dividing cells with low metabolic activity, named persisters, are tolerant to antibiotic treatment and are the principal cause of recalcitrant and resistant infections, including skin infections. Antimicrobial peptides (AMPs) hold promise as new anti-infective agents to treat such infections. Here for the first time, we investigated the activity of the frog-skin AMP temporin G (TG) against preformed S. aureus biofilm including persisters, as well as its efficacy in combination with tobramycin, in inhibiting S. aureus growth. TG was found to provoke ~50 to 100% reduction of biofilm viability in the concentration range from 12.5 to 100 µM vs ATCC and clinical isolates and to be active against persister cells (about 70–80% killing at 50–100 µM). Notably, sub-inhibitory concentrations of TG in combination with tobramycin were able to significantly reduce S. aureus growth, potentiating the antibiotic power. No critical cytotoxicity was detected when TG was tested in vitro up to 100 µM against human keratinocytes, confirming its safety profile for the development of a new potential anti-infective drug, especially for treatment of bacterial skin infections.     

SBBR反应器中耐冷微生物的驯化与识别

以模拟生活污水为处理对象，三种不同填料为生物膜载体，运行 A、B、C三个SBBR，通过逐步降温（25、20、15、10、6和5℃）培养驯化处理低温污水的生物膜。运行251周期，三个反应器COD出水浓度均低于50 mg/L；B反应器对氨氮去除效果优于A和C。特别在低温5℃时，A、B和C三个反应器氨氮平均出水浓度分别为14.1、3.79和14.1 mg/L。高通量测序结果显示，5℃时，三个反应器中均以降解有机物的微生物为主；只有B反应器中驯化富集出了嗜冷硝化菌（Candidatus Nitrotoga）；而其他有硝化作用的菌属在B反应器中的丰度也高于A和C反应器；不利于脱氮的类固氮菌（Elstera）在A和C反应器中丰度较高。此结果可从微生物角度解释三个反应器处理效果的差异性，表明B反应器处理低温污水有显著优势，其所用填料可结合低温菌进一步开发利用。     

船用一体化生物活性炭装置反冲洗技术条件优化

反冲洗是保证生物活性炭装置正常运行的重要因素。以船用污水深度处理装置一体化生物活性炭为研究对象,探究反冲洗对于装置污染物去除能力恢复的影响,得出了装置恢复时间;选取9种反冲洗条件,考察装置反洗后8 h后,装置污染物去除能力的变化。结果表明:污染物去除率随着反冲洗时间推移先降低后逐步升高,8 h后趋于稳定,此时COD去除率为60%,氨氮去除率为10%。对比不同反冲洗条件下对于装置性能的影响分析,反洗强度控制在5.0 L/(m2·s),反洗时间为7 min时为较优反冲洗条件,COD去除率达到79.8%,氨氮去除率为5.3%。