Influences and mechanisms of nanofullerene on the horizontal transfer of plasmid-encoded antibiotic resistance genes between E. coli strains

• Sub-inhibitory levels of nC₆₀ promote conjugative transfer of ARGs. • nC₆₀ can induce ROS generation, oxidative stress and SOS response. • nC₆₀ can increase cell membrane permeability and alter gene expression. • Results provide evidence of nC₆₀ promoting antibiotic resistance dissemination. The spread and development of antibiotic resistance globally have led to severe public health problems. It has been shown that some non-antibiotic substances can also promote the diffusion and spread of antibiotic resistance genes (ARGs). Nanofullerene (nC₆₀) is a type of nanomaterial widely used around the world, and some studies have discovered both the biological toxicity and environmental toxicity of nC₆₀. In this study, cellular and molecular biology techniques were employed to investigate the influences of nC₆₀ at sub-minimum inhibitory concentrations (sub-MICs) on the conjugation of ARGs between the E. coli strains. Compared with the control group, nC₆₀ significantly increased the conjugation rates of ARGs by 1.32‒10.82 folds within the concentration range of 7.03‒1800 mg/L. This study further explored the mechanism of this phenomenon, finding that sub-MICs of nC₆₀ could induce the production of reactive oxygen species (ROS), trigger SOS-response and oxidative stress, affect the expression of outer membrane proteins (OMPs) genes, increase membrane permeability, and thus promote the occurrence of conjugation. This research enriches our understanding of the environmental toxicity of nC₆₀, raises our risk awareness toward nC₆₀, and may promote the more rational employment of nC₆₀ materials.     

镉对大鼠肾皮质结合反应的影响

通过体内和体外试验分别研究了镉对肾皮质微粒体谷胱甘肽转移酶(GST)及游离肾近曲小管细胞中葡萄糖醛酸(GA)结合反应的影响.结果表明:(1)体外试验中,在10~(-7)—10~(-3)mol/L浓度范围内,镉可抑制肾皮质微粒休GST活力;(2)体内腹腔注射CdCl_2(2.4mg/kg,连续3天)后,大鼠肾皮质GST活力无明显改变;(3)按同样方式腹腔注射CdCl_2后,可降低游离肾近曲小管细胞中GA和7-羟基香豆素(7-HC)的结合反应;(4)体外试验中,低浓度镉(10~(-7)mol/L)可使肾皮质细胞中GA和7-HC的结合反应增强,而在10~(-6)—10~(-2)mol/L浓度范围内,镉可降低GA和7-HC的结合反应.     

Herbicide promotes the conjugative transfer of multi-resistance genes by facilitating cellular contact and plasmid transfer

The global dissemination of antibiotic resistance genes (ARGs), especially via plasmid-mediated horizontal transfer, is becoming a pervasive health threat. While our previous study found that herbicides can accelerate the horizontal gene transfer (HGT) of ARGs in soil bacteria, the underlying mechanisms by which herbicides promote the HGT of ARGs across and within bacterial genera are still unclear. Here, the underlying mechanism associated with herbicide-promoted HGT was analyzed by detecting intracellular reactive oxygen species (ROS) production, extracellular polymeric substance composition, cell membrane integrity and proton motive force combined with genome-wide RNA sequencing. Exposure to herbicides induced a series of the above bacterial responses to promote HGT except for the ROS response, including compact cell-to-cell contact by enhancing pilus-encoded gene expression and decreasing cell surface charge, increasing cell membrane permeability, and enhancing the proton motive force, providing additional power for DNA uptake. This study provides a mechanistic understanding of the risk of bacterial resistance spread promoted by herbicides, which elucidates a new perspective on nonantibiotic agrochemical acceleration of the HGT of ARGs.     

Impacts of organic carbon availability and recipient bacteria characteristics on the potential for TOL plasmid genetic bioaugmentation in soil slurries

The effectiveness of genetic bioaugmentation relies on efficient plasmid transfer between donor and recipient cells as well as the plasmid’s phenotype in the recipient cell. In the present study, the effects of varying organic carbon substrates, initial recipient-to-donor cell density ratios, and mixtures of known recipient bacterial strains on the conjugation and function of a TOL plasmid were tested in sterile soil slurry batch reactors. The presence of soil organic carbon was sufficient in ensuring TOL plasmid transconjugant occurrence (up to 2.1 ± 0.5%) for most recipient strains in soil slurry batch mating experiments. The addition of glucose had limited effects on transconjugant occurrence; however, glucose amendment increased the specific toluene degradation rates of some Enterobacteriaceae transconjugants in soil slurry. Initial cell density ratios and mixtures of recipient strains had smaller impacts on plasmid conjugation and resulting phenotype functionality. These observations suggest that genetic bioaugmentation may be improved by minimal altering of environmental conditions.