Cousins C M, Melin J R, Venables W A, Coakley W T
School of Biosciences, Cardiff University, UK.
Bioseparation. 2000;9(6):343-9. doi: 10.1023/a:1011181431742.
Cells aggregate and can be recovered from suspension when exposed to an ultrasonic standing wave field. The acoustic force on individual cells in a standing wave decreases with particle volume. A plane ultrasonic field generated by a transducer driven at 3.3 MHz was used here to investigate the removal of Escherischia coli, cells with dimensions of the order of 1.0 microm, from batch suspension by sedimentation over a range of concentrations (10(3) to 10(10) cells ml(-1)). Cell removal efficiencies greater than 90% were achieved at initial concentrations of 10(10) cells ml(-1). Removal efficiencies decreased gradually to zero, as initial bacterial concentration was reduced to 10(7) cells ml(-1). It was found that, when low concentrations of E. coli (10(3) to 10(5) cells ml(-1)) were added to suspensions of larger particles (i.e. yeast cells) that were of sufficient concentration to form aggregates in the sound field, E. coli could be harvested to an efficiency of 40%. The results imply that the E. coli became trapped and sediment with aggregates of larger particles. Some strains of bacteria are capable of DNA transfer by conjugation. The transfer rate of E. coli RP4 plasmid is order of magnitudes greater when conjugation occurs on solid medium rather than in liquid suspension. We have investigated whether the conjugation rate would also be higher in ultrasonically induced E. coli clumps than in free suspension. The donor strain was mixed with a recipient strain of E. coli, then sonicated in a capillary at 4.6 MHz in a tubular transducer for 5 min. The bacteria aggregated successfully. Results showed a three-fold increase in the rate of conjugation compared to a liquid mating control.
当暴露于超声驻波场时,细胞会聚集并可从悬浮液中回收。驻波中单个细胞上的声学力随颗粒体积减小。此处使用由3.3 MHz驱动的换能器产生的平面超声场,来研究在一系列浓度(10³至10¹⁰个细胞/毫升)下,通过沉降从分批悬浮液中去除尺寸约为1.0微米的大肠杆菌细胞的情况。在初始浓度为10¹⁰个细胞/毫升时,细胞去除效率达到90%以上。随着初始细菌浓度降至10⁷个细胞/毫升,去除效率逐渐降至零。研究发现,当将低浓度的大肠杆菌(10³至10⁵个细胞/毫升)添加到足够浓度以在声场中形成聚集体的较大颗粒(即酵母细胞)的悬浮液中时,大肠杆菌的收获效率可达40%。结果表明,大肠杆菌被困住并与较大颗粒的聚集体一起沉降。一些细菌菌株能够通过接合进行DNA转移。当接合在固体培养基上而非液体悬浮液中发生时,大肠杆菌RP4质粒的转移速率要高几个数量级。我们研究了在超声诱导的大肠杆菌团块中接合率是否也会高于游离悬浮液中的接合率。将供体菌株与大肠杆菌受体菌株混合,然后在管状换能器中于4.6 MHz下在毛细管中超声处理5分钟。细菌成功聚集。结果表明,与液体交配对照相比,接合率提高了三倍。