1.Release of DNA from Pseudomonas aeruginosa in vitro during spontaneous growth and treatment with ciprofloxacin.
Dai-Zhi PENG ; Charles H GUYMON ; Albert T MCMANUS ; Guang-Xia XIAO
Chinese Journal of Surgery 2005;43(3):178-181
OBJECTIVETo observe the DNA release from Pseudomonas aeruginosa (P. aeruginosa) during spontaneous growth and exposure to different concentrations of ciprofloxacin (Cipro) in vitro.
METHODSThe P. aeruginosa 1244 strain (ATCC 27317) was selected because it was sensitive to Cipro in vitro. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of Cipro against this strain were determined, respectively. Different concentrations of Cipro were cultured with this strain at 37 degrees C for 4 h and 24 h. The samples of culture supernatant were filtered and electrophoresed in 1.8% agarose with SYBR Gold stain. Then the images of the gel sheets were photographed.
RESULTSThe MIC and MBC of Cipro were 0.25 - 0.5 mg/L. The free bacterial DNA in 4 h culture samples with or without Cipro could not be detected by this method. The certain amount of free bacterial DNA molecules in 24 h culture samples without antibiotic appeared at the two zones whose molecular weights were more than 2000 bp and less than 100 bp. The large amount of free bacterial DNA molecules showed at three zones in 24 h culture samples with Cipro when its concentrations were much lower than its MIC. In terms of DNA molecular weight, the first two zones were above 2000 bp, and the third zone was below 100 bp. There was no detectable DNA release from bacteria in 24 h culture samples when Cipro was at or above its MIC.
CONCLUSIONSThe certain amount of bacterial DNA were released from P. aeruginosa in the spontaneous growth. Different concentrations of Cipro had quite differential effects on the DNA release from P. aeruginosa in quantities and molecular weights in vitro.
Anti-Infective Agents ; administration & dosage ; pharmacology ; Ciprofloxacin ; administration & dosage ; pharmacology ; DNA, Bacterial ; metabolism ; Dose-Response Relationship, Drug ; In Vitro Techniques ; Microbial Sensitivity Tests ; Pseudomonas aeruginosa ; drug effects ; metabolism
2.Preliminary study on the release of DNA from Pseudomona aeruginosa induced by piperacillin/tazobactam in vitro.
Dai-zhi PENG ; Charles H GUYMON ; Albert T MCMANUS ; Guang-xia XIAO
Chinese Journal of Burns 2005;21(2):93-96
OBJECTIVETo observe the release of DNA from Pseudomonas aeruginosa (P. aeruginosa) induced by different concentrations of piperacillin/tazobactam (Piper) in vitro.
METHODSThe minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of Piper against 1244 strain (ATCC 27317) of P. aeruginosa were determined, respectively. This strain of P. aeruginosa was separately cultured with Piper in different concentrations at 37 degrees C for 4 h and 24 h. The samples of cultural supernatant were filtered and electrophoresis was conducted in 1.8% agarose with SYBR Gold stain. Then the images of the gel sheets were photographed.
RESULTSThis strain of P. aeruginosa was sensitive to Piper. The bacterial DNA was not detected in 4-h cultured P. aeruginosa either with or without Piper by this method. The bacterial DNA molecules could be detected in 24 h samples in cultures without Piper, and they were displayed in two zones of molecular weight over 2000 base pairs (bp) and lower than 100 bp. Similar results were observed when the MIC of piper (0.002, 0.004 g/L) were under the MIC measured at the 3rd time (0.008 g/L), but there was much more bacterial DNA with molecular weight lower than 100 bp. When Piper concentration was higher than its MIC, only smaller quantities of bacterial DNA in the area with molecular weight lower than 400 bp could be detected in 24-h culture samples.
CONCLUSIONA certain amount of bacterial DNA was released from P. aeruginosa under its natural growth circumstance. Different concentrations of Piper showed different effects on DNA release, in regard to its quantity and molecular weight, from P. aeruginosa cultures.
Anti-Bacterial Agents ; pharmacology ; DNA, Bacterial ; metabolism ; Penicillanic Acid ; analogs & derivatives ; pharmacology ; Piperacillin ; pharmacology ; Pseudomonas aeruginosa ; drug effects ; metabolism