Department of Civil and Environmental Engineering, Duke University, Durham, North Carolina 27708, United States.
Environ Sci Technol. 2022 Nov 1;56(21):15044-15053. doi: 10.1021/acs.est.2c02878. Epub 2022 Jul 19.
Extracellular antibiotic resistance genes (eARGs) are widespread in the environment and can genetically transform bacteria. This work examined the role of environmentally relevant nanoparticles (NPs) in regulating eARG bioavailability. eARGs extracted from antibiotic-resistant were incubated with nonresistant recipient cells. In the mixture, particle type (either humic acid coated nanoparticles (HASNPs) or their micron-sized counterpart (HASPs)), DNase I concentration, and eARG type were systematically varied. Transformants were counted on selective media. Particles decreased bacterial growth and eARG bioavailability in systems without nuclease. When DNase I was present (≥5 μg/mL), particles increased transformation via chromosomal (but not plasmid-borne) eARGs. HASNPs increased transformation more than HASPs, indicating that the smaller nanoparticle with greater surface area per volume is more effective in increasing eARG bioavailability. These results were also modeled via particle aggregation theory, which represented eARG-bacteria interactions as transport leading to collision, followed by attachment. Using attachment efficiency as a fitting factor, the model predicted transformant concentrations within 35% of experimental data. These results confirm the ability of NPs to increase eARG bioavailability and suggest that particle aggregation theory may be a simplified and suitable framework to broadly predict eARG uptake.
细胞外抗生素耐药基因(eARGs)广泛存在于环境中,并能使细菌发生基因转化。本研究考察了环境相关纳米颗粒(NPs)在调节 eARG 生物可利用性方面的作用。从抗生素耐药菌中提取的 eARGs 与非耐药受体菌细胞共孵育。在混合物中,系统改变了颗粒类型(例如,腐殖酸包覆纳米颗粒(HASNPs)或其微米级对应物(HASPs))、DNase I 浓度和 eARG 类型。在选择性培养基上计数转化体。在没有核酸酶的系统中,颗粒会降低细菌生长和 eARG 的生物可利用性。当存在 DNase I(≥5μg/mL)时,颗粒通过染色体(而非质粒携带)eARGs 增加转化。HASNPs 比 HASPs 更能增加转化,这表明比表面积与体积比更大的较小纳米颗粒更能有效提高 eARG 的生物可利用性。这些结果还通过颗粒聚集理论进行了建模,该理论将 eARG-细菌相互作用表示为导致碰撞的传输,随后是附着。使用附着效率作为拟合因子,该模型预测的转化体浓度与实验数据相差 35%以内。这些结果证实了 NPs 增加 eARG 生物可利用性的能力,并表明颗粒聚集理论可能是一种简化且合适的框架,可广泛预测 eARG 的摄取。