Departamento de Ingeniería Química y Alimentos, Universidad de las Américas Puebla, Cholula, Puebla 72820, México.
J Food Prot. 2009 Nov;72(11):2301-7. doi: 10.4315/0362-028x-72.11.2301.
Escherichia coli ATCC 35218 growth response was evaluated after repetitive cultivation in stepwise increasing antimicrobial agent concentrations (potassium sorbate or sodium benzoate) to observe its adaptation process to high weak-acid concentrations. The effect of antimicrobial (potassium sorbate or sodium benzoate) concentration (0 to 7,000 ppm) was tested using laboratory media. Cells adapted at 1,000 ppm were inoculated in media containing the same concentration of the antimicrobial; after that, cells were transferred to media containing a higher concentration, followed by repetitive cultivations. In every case, viable cells were determined by surface plating every hour up to 48 h. Logarithmic representations of survival or growing fraction were modeled using the Gompertz equation. Adapted and nonadapted cells were analyzed for plasmid presence as well as phosphofructokinase and succinate dehydrogenase activity. Bacterial growth was observed after adaptation processes in media formulated up to 7,000 ppm of potassium sorbate or sodium benzoate. Analyses of variance demonstrated that no significant difference (P > 0.05) in lag time or growth rate was observed among adapted cells cultured in media containing the studied concentrations for each of the antimicrobials tested. These results suggest that E. coli can be adapted to high weak-acid concentrations if the exposure is performed under sublethal conditions. Furthermore, there was demonstrated inhibition of the enzymes phosphofructokinase and succinate dehydrogenase by action of sodium benzoate and potassium sorbate, respectively. E. coli adaptation to antimicrobial agents was not related to plasmid presence but appears to be due to other action mechanisms.
评估了大肠杆菌 ATCC 35218 在逐步增加抗菌剂浓度(山梨酸钾或苯甲酸钠)的重复培养中的生长反应,以观察其适应高弱酸浓度的过程。使用实验室培养基测试了抗菌剂(山梨酸钾或苯甲酸钠)浓度(0 至 7000 ppm)的影响。将适应 1000 ppm 的细胞接种到含有相同浓度抗菌剂的培养基中;之后,将细胞转移到含有更高浓度的培养基中,然后进行重复培养。在每种情况下,通过表面平板法每小时测定存活细胞数,直至 48 小时。使用 Gompertz 方程对存活或生长分数的对数表示进行建模。分析了适应和不适应细胞的质粒存在情况以及磷酸果糖激酶和琥珀酸脱氢酶活性。在含有高达 7000 ppm 山梨酸钾或苯甲酸钠的培养基中进行适应过程后,观察到细菌生长。方差分析表明,在含有测试抗菌剂每种浓度的培养基中培养的适应细胞的滞后时间或生长率之间没有显著差异(P > 0.05)。这些结果表明,如果暴露在亚致死条件下,大肠杆菌可以适应高弱酸浓度。此外,山梨酸钾和苯甲酸钠分别抑制了磷酸果糖激酶和琥珀酸脱氢酶的活性。大肠杆菌对抗菌剂的适应性与质粒的存在无关,但似乎是由于其他作用机制。