[Study on the transfer mechanism and destructive law of the bacterial antibiotic resistance genes disinfected by chlorination]

Zhonghua Yu Fang Yi Xue Za Zhi. 2018 Sep 6;52(9):892-897. doi: 10.3760/cma.j.issn.0253-9624.2018.09.005.
[Article in Chinese]

Abstract

Objective: To study the response of Escherichia coli (E. coli) HB101 (plasmid pUC19) and its carried antibiotic resistance genes to the process of cholorination under different environmental conditions. Methods: The E. coli strain was reacted with sodium hypochlorite at the concentration of 0.5, 0.75, 1.00, and 0.55 mg/L, then the residual chlorine and the colonies were detected at the 0.25, 1, 2, 5, 10, 20, and 30 min of the reaction, respectively. The first order disinfection kinetic model and EFH model were used to evaluate the inactivation effect of E. coli (plasmid pUC19) treated by sodium hypochlorite, while the plasmid pUC19 and antibiotic resistance gene amp(r) were detected by PCR method. Besides, the logarithm of Ct (residual chlorine in t) under different concentration were calculated. Results: The temperature and pH value played important roles on the inactivation of E. coli and elimination of plasmid pUC19 and amp(r) under the function of sodium hypochlorite. The Ct value needed for 5-log of E.coli HB101(pUC19) inactivation at 4, 20, 36 ℃ was 11.92, 10.28, 7.67, respectively, and when the pH was in 6.0, 7.0, 8.0, with chloride concentration were 0.75, 0.70, 0.55 mg/L, the Ct value needed for reached to 6.68, 10.28, 15.73 min·mg/L. At pH 7.2 condition, when the temperature was 4, 20, 36 ℃, and chloride concentration were 9, 5, 3 mg/L.The required Ct values to completely destroy the transformation function of free antibiotic resistant plasmids were 36.11, 34.17,16.09 min·mg/L. Sodium hypochlorite disinfection can release free ampr gene and even the transformed plasmid pUC19, and pollute the water body. Only when the Ct value reached 903.03 min·mg/L, the complete ampr gene can be destroyed which was far more exceed the bacterial lethal Ct value. Conclusion: Even if all the antibiotic resistant bacteria were inactivated, the antibiotic resistant plasmids or genes might still maintain complete with the transformable function, which may result in new potential risks of waterborne diseases.

目的:研究氯化消毒在不同环境条件下对抗生素抗性细菌大肠埃希菌(E. coli)HB101(pUC19质粒)的灭活规律以及对其抗生素抗性基因的转移和破坏机制。 方法:E. coli菌株与0.5、0.75、1.00、0.55 mg/L次氯酸钠作用后,分别在0.25、1、2、5、10、20、30 min的处理时间进行余氯测定、菌落计数,并采用消毒一级动力学模型和灭活动力学EFH模型进行拟合,获得E. coli的灭活曲线,同时,采用PCR技术进行抗生素抗性质粒pUC19质粒及其氨苄青霉素抗性基因(amp(r))的检测,利用细菌转化试验评价抗生素抗性质粒的功能,计算达到不同的细胞杀灭对数值时的Ct值(t时刻的余氯浓度)。 结果:消毒温度和pH值对水中E. coli的杀灭、pUC19质粒和amp(r)的破坏具有显著影响,当消毒温度为4、20、36 ℃时,细胞消灭对数值达到5时所需Ct值分别为11.92、10.28、7.67 min·mg/L。同时,当消毒pH分别为6.0、7.2、8.0,氯浓度分别为0.75、0.70、0.55 mg/L时,细胞消灭对数值达到5时所需Ct值分别为6.68、10.28、15.73 min·mg/L。pH 7.2且消毒温度为4、20、36 ℃,氯浓度分别为9、5、3 mg/L时,为完全破坏细胞外游离抗生素抗性质粒的转化功能,消毒所需Ct值分别为36.11、34.17、16.09 min·mg/L。次氯酸钠消毒会导致游离amp(r)基因甚至具有转化功能的完整质粒pUC19质粒释放,并污染水体,只有当Ct值达到903.03 min·mg/L之后,完整的amp(r)基因才能被消除,远远超过细菌致死Ct值。 结论:次氯酸钠消毒过程中,抗生素抗性细菌E. coli全部被破坏,但其携带抗生素抗性的DNA片段(质粒和基因)仍可能保持完整性且具有转化功能。.

Keywords: Antibiotic resistance genes; Antibiotics; Disinfection; Water.

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Disinfection / methods
  • Drug Resistance, Bacterial / genetics*
  • Escherichia coli / drug effects
  • Escherichia coli / genetics*
  • Halogenation
  • Humans

Substances

  • Anti-Bacterial Agents